TWI856340B - Heterocyclic compounds as immunomodulators of pd-l1 interactions - Google Patents

Abstract

PD-L1 inhibitors of various compound formulas, both generically and specifically are disclosed. Methods of making such PD-L1 inhibitor compounds are disclosed, both generically and specifically. Methods of using such PD-L1 inhibitor compounds singly or in combination with additional agents and compositions of such PD-L1 inhibitor compounds for the treatment of cancer and other conditions are disclosed.

Description

Heterocyclic compounds as PD-L1-interacting immunomodulators

The present application generally relates to chemical compounds, and more particularly to compounds that modulate the biological activity of PD-L1 protein.

Programmed death-ligand 1 (PD-L1) is a protein that plays an important role in the adaptive arm of the immune system. Normally, the immune system is self-regulated and responds to antigens associated with immune system activation through exogenous or endogenous danger signals. As a result, the replication, expansion and spread of antigen-specific CD8+ T cells and/or CD4+ helper cells occur. Binding of PD-L1 to the inhibitory checkpoint molecule PD-1 on T cells transmits an inhibitory signal that reduces the proliferation of antigen-specific T cells in lymph nodes and reduces apoptosis in regulatory T cells (anti-inflammatory, suppressive T cells).

Therefore, molecules that can modulate PD-L1 activity can be widely used in the treatment of various disease states.

One aspect of the present application relates to a compound having a general structure as shown in formula (I): (I)

or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate or tautomer thereof, wherein:

A and B are each independently selected from halogen, cyano, -N ₃ , alkyl and substituted alkyl, amine, alkylamine, alkoxy;

Z ₁ is -CR ¹ = or -N =;

Z ₂ is -CR ² =;

Z ₃ is -CR ³ = or -N =;

Z ₄ is -CR ⁴ = or -N =;

Z ₅ is -CR ⁵ =;

Z ₆ is -CR ⁶ = or -N =;

^R1 and ^R4 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

^R2 and ^R5 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

^R3 and ^R6 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

_Y1 and _Y2 are independently -C( ^R7 )( ^R8 )-, ^-CR9 =, ^-NR10- , -O- or -S-;

_X1 and _X2 are each independently -C( ^R11 )( ^R12 )-, -N=, ^-NR13- , -S- or -O-;

R ⁷ , R ⁸ , R ⁹ , R ¹¹ and R ¹² are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

R ¹⁰ and R ¹³ are each independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, alkylamine, alkoxy;

_L1 and _L2 are each an alkyl group, a substituted alkyl group or a heteroatom chain containing m atoms between ring 3 and _W1 and between ring 6 and _W2 , wherein m=0, 1, 2, 3, 4, 5 or 6; when m is 0, _W1 or _W2 is directly connected to the corresponding nitrogen in ring 3 or ring 6, respectively;

_W1 and _W2 are each independently hydrogen, a five-membered heterocyclic ring or a substituted five-membered heterocyclic ring, a six-membered heterocyclic ring or a substituted six-membered heterocyclic ring, a carboxyalkyl group or a substituted carboxyalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an amino acid ester, an amino acid amide, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid.

Another aspect of the present application relates to a method for treating a disease or disease state associated with the interaction between PD-L1 and PD-1 in an individual, the method comprising the following steps: administering to the individual an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate or tautomer thereof.

Another aspect of the present application relates to a process for preparing a compound of formula (I).

Reference will be made in detail to certain aspects and exemplary embodiments of the present application, examples of which are shown in the attached structures and drawings. Various aspects of the present application, including methods, materials and examples, will be described in conjunction with the exemplary embodiments, such description is non-restrictive, and the scope of the present application is intended to include all equivalents, substitutes and modifications that are generally known or incorporated herein. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art to which the present application belongs. Those of ordinary skill in the art will recognize many technologies and materials that are similar or equivalent to the technologies and materials described herein, which can be used to implement various aspects and embodiments of the present application. The aspects and embodiments described in the present application are not limited to the methods and materials described.

As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

In this article, a range can be expressed as from "about" a specific value and/or to "about" another specific value. When such a range is expressed, another embodiment includes from a specific value and/or to another specific value. Similarly, when a value is expressed as an approximation by using the antecedent "about", it is understood that the specific value forms another embodiment. It can be further understood that the endpoints of each range are significant in relation to the other endpoint and independently of the other endpoint. It can also be understood that multiple values are disclosed herein, and in addition to the value itself, each value is also disclosed herein as "about" the specific value. For example, if the value "10" is disclosed, "about 10" is also disclosed. It can also be understood that when a value "less than or equal to" the value is disclosed, "greater than or equal to the value" and the possible range between the values are also disclosed, as appropriately understood by those with ordinary knowledge in the art. For example, if the value "10" is disclosed, then "less than or equal to 10" and "greater than or equal to 10" are also disclosed.

At various places in this specification, certain features of compounds are disclosed in groups or in ranges. It is specifically intended that such disclosure includes every individual subcombination of the members of such groups and ranges.

The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as mirror isomers and non-mirror isomers, are intended. The compounds of the present application containing asymmetric substituted carbon atoms can be separated in optically active or racemic forms. How to prepare optically active forms from optically inactive starting materials, for example, by the resolution of a racemic mixture or by stereoselective synthesis is known in the art. Many geometric isomers of alkenes, C=N double bonds, etc. can also be present in the compounds described herein, and all such stable isomers are considered in the present application. The cis and trans geometric isomers of the compounds of the present application are described and can be separated into mixtures of isomers or separated isomeric forms.

The resolution of the racemic mixture of the compound can be carried out by any of a variety of methods known in the art. One method includes partial recrystallization using a chiral resolving acid, which is an optically active, salt-forming organic acid. Suitable resolving agents for the partial recrystallization method are, for example, optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, apple acid, lactic acid, or various optically active camphorsulfonic acids such as b-camphorsulfonic acid. Other resolving agents suitable for the partial crystallization method include stereoisomerically pure forms of a-methylbenzylamine (e.g., S and R forms or non-stereoisomerically pure forms), 2-phenylglycinol, phenylpropanolamine, ephedrine, TV-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

The racemic mixture can also be resolved by eluting on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). One of ordinary skill in the art will be able to determine the appropriate elution solvent composition.

In compounds with more than one chiral center, each chiral center in the compound may independently be (R) or (S), unless otherwise indicated.

The compounds of the present application also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond with the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of proton-isotropic tautomers include keto-enol pairs, amide-imidic acid pairs, lactamide-lactimide pairs, enamine-imine pairs, and cyclic forms in which the proton can occupy two or more positions of the heterocyclic system, for example, 1H- and 3/f-imidazole, 1H-, 2- and 4H-1,2,4-triazole, \H- and 2H-isoindole and 1H- and 2//-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

The compounds of the present application can also include all isotopes of atoms produced in the intermediates or final compounds. Isotopes include those atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present application can be replaced or substituted with isotopes of atoms of natural or unnatural abundance. In certain embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in the disclosed compounds can be replaced or substituted with deuterium. In certain embodiments, the compound includes two or more deuterium atoms. I. Definitions

As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. The term is also intended to coreference the compounds of the present application, regardless of how they are prepared, e.g., synthetically, by biological processes (e.g., metabolic or enzymatic conversion), or a combination thereof.

All compounds and their pharmaceutically acceptable salts can exist together with other substances such as water and solvents (e.g., hydrates and solvates), or can be separated. When in a solid state, the compounds and their salts described herein can exist in various forms, and can be in the form of solvates, including hydrates. The compound can be in any solid form, such as a polymorph or a solvate, so unless otherwise expressly indicated, reference to the compound and its salt in the specification should be understood to cover the compound in any solid form.

In certain embodiments, the compounds of the present invention or their salts are substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which the compound is formed or detected. Partial isolation can include, for example, a composition enriched with the compounds of the present invention.

Substantial isolation can include compositions containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compound or salt thereof.

The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio.

As used herein, the terms "ambient temperature" and "room temperature" are understood in the art and generally refer to a temperature, such as a reaction temperature, that is approximately the temperature of the room in which the reaction is conducted, for example, a temperature of about 20°C to about 30°C.

The present application also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salt" refers to a derivative of a disclosed compound in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of alkaline residues such as amines; alkaline or organic salts of acidic residues such as carboxylic acids, etc. The pharmaceutically acceptable salts of the present application include, for example, non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present application can be synthesized from a parent compound containing an alkaline or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol or butanol) or acetonitrile (MeCN) are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci, 1977, 66(1), 1-19, and in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In certain embodiments, the compounds described herein include N-oxide forms.

The terms "subject" or "patient" are used interchangeably to refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and preferably humans.

The phrase "therapeutically effective amount" means that amount of an active compound or agent that elicits the biological or medicinal response that is being sought by the researcher, veterinarian, physician or other clinician.

As used herein, the term "treating" refers to one or more of the following: (1) inhibiting a disease; e.g., inhibiting the pathology or symptoms of a disease, disease state or disorder in a subject that is experiencing or displaying the disease, disease state or disorder (i.e., arresting further progression of the pathology and/or symptoms); and (2) ameliorating a disease; e.g., ameliorating a disease, disease state or disorder, such as reducing the severity of the disease, in a subject that is experiencing or displaying the disease, disease state or disorder (i.e., reversing the pathology and/or symptoms).

The term "solvate" refers to a compound formed by the interaction of a solvent with an EPI, a metabolite or a salt thereof. Suitable solvates are pharmaceutically acceptable solvates including hydrates.

The term "substituted" or "optionally substituted" as used in the present invention means that one or more hydrogen atoms of the group referred to by the term "substituted" or "optionally substituted" are replaced by one of the substituents independently selected from the following: lower alkyl, lower aryl, lower aralkyl, lower cycloalkyl, lower heterocycloalkyl, hydroxy, lower alkoxy, lower aryloxy, perhalogenated alkoxy, aralkyloxy, lower heteroaryl, lower heteroaryloxy, lower heteroarylalkyl, lower heteroaralkyloxy, azido, amino, halogen, lower alkylthio, oxo, lower acylalkyl, lower carboxyl ester, carboxyl, amido, nitro, Lower acyloxy, lower aminoalkyl, lower alkylaminoaryl, lower alkylaryl, lower alkylaminoalkyl, lower alkoxyaryl, lower arylamino, lower arylalkylamino, sulfonyl, lower amidoalkylaryl, lower amidoaryl, lower hydroxyalkyl, lower haloalkyl, lower alkylaminoalkylcarboxy, lower aminoamidoalkyl, cyano, lower alkoxyalkyl, lower perhaloalkyl and lower arylalkoxyalkyl, provided that the normal valence of the atom being substituted is not exceeded and the substitution results in a stable chemical compound, i.e., a compound sufficiently stable to be isolated from the reaction mixture.

The term "alkyl" refers to a straight or branched chain or cyclic hydrocarbon group having only a single carbon-carbon bond. Representative examples include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl and cyclohexyl, all of which may be optionally substituted.

The term "aryl" refers to aromatic groups having 5 to 14 ring atoms and at least one ring having a conjugated π electron system, and includes carbocyclic aryl, heterocyclic aryl and biaryl, all of which may be optionally substituted.

Carbocyclic aryl is a group having 6 to 14 ring atoms, wherein the ring atoms on the aromatic ring are carbon atoms.Carbocyclic aryl includes monocyclic carbocyclic aryl and polycyclic or fused compounds, such as optionally substituted naphthyl.

Heterocyclic aryl or heteroaryl is a group having 5 to 14 ring atoms, of which 1 to 4 heteroatoms are ring atoms in an aromatic ring and the remaining ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, nitrogen and selenium. Suitable heteroaryl groups include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolyl, pyridyl-N-oxide, pyrimidinyl, pyrazinyl, imidazolyl and the like, all of which are optionally substituted.

The term "diaryl" refers to aryl groups having 5 to 14 atoms containing more than one aromatic ring, including fused ring systems and aryl groups substituted with other aryl groups. Such groups may be optionally substituted. Suitable diaryl groups include naphthyl and biphenyl.

The terms "substituted aryl" and "substituted heteroaryl" refer to aryl and heteroaryl groups substituted with 1 to 3 substituents selected from lower alkyl, lower alkoxy, lower perhalogenated alkyl, halogen, hydroxyl and amino.

The term "aralkyl" refers to an alkylene group substituted by an aryl group. Suitable aralkyl groups include benzyl, picolyl, etc., and may be optionally substituted.

The term "heteroarylalkyl" refers to an alkylene group substituted with a heteroaryl group.

The term "alkylaryl" refers to an aryl group substituted by an alkyl group. "Lower alkylaryl" refers to those groups in which the alkyl group is a lower alkyl group.

The term "lower" as used herein in connection with an organic group or compound, respectively, refers to 6 carbon atoms or less. Such groups may be linear, branched or cyclic.

The term "higher" as used herein in connection with an organic group or compound, respectively, refers to 7 or more carbon atoms. Such groups may be linear, branched or cyclic.

The term "cyclic alkyl" or "cycloalkyl" refers to a cyclic alkyl group having from 3 to 10 carbon atoms, and in one aspect from 3 to 6 carbon atoms. Suitable cyclic groups include norbornyl and cyclopropyl. Such groups may be substituted.

The term "heterocyclic", "heterocyclic alkyl" or "heterocycloalkyl" refers to a cyclic group having from 3 to 10 atoms, and in one aspect a cyclic group having from 3 to 6 atoms containing at least one heteroatom, in another aspect 1 to 3 heteroatoms. Suitable heteroatoms include oxygen, sulfur and nitrogen. The heterocyclic group may be attached via nitrogen or carbon atoms in the ring. Heterocycloalkyl groups include unsaturated cyclic, fused cyclic and spirocyclic groups. Suitable heterocyclic groups include pyrrolidinyl, morpholinyl, morpholinylethyl and pyridyl.

The terms "arylamino" (a) and "aralkylamino" (b) refer to the radical -NRR', wherein (a) R is aryl and R' is hydrogen, alkyl, aralkyl, heterocycloalkyl or aryl, and (b) R' is aralkyl and R' is hydrogen, aralkyl, aryl, alkyl or heterocycloalkyl, respectively.

The term "acyl" refers to -C(O)-R, where R is alkyl, heterocycloalkyl or aryl.

The term "carboxy ester" refers to -C(O)-OR, where R is alkyl, aryl, aralkyl, cycloalkyl or heterocycloalkyl, all optionally substituted.

The term "carboxy" refers to -C(O)-OH.

The term "oxo" refers to =0 in an alkyl or heterocycloalkyl group.

The term "amino" refers to -NRR', wherein R and R' are independently selected from hydrogen, alkyl, aryl, aralkyl and heterocycloalkyl, all of which except H are optionally substituted; and R and R' can form a cyclic ring system.

The term "amido" refers to -C(O) _NR2 , wherein each R is independently hydrogen or alkyl.

The term "sulfonamido" refers to -S(=O) ₂ R ₂ , wherein each R is independently hydrogen or alkyl.

The term "halogen" refers to -F, -Cl, -Br and -I.

The term "alkylaminoalkylcarboxy" refers to the group alkyl-NR-alk-C(O)-O-, where "alk" is alkylene and R is H or lower alkyl.

The term "sulfonyl" refers to _-SO2R , where R is H, alkyl, aryl, aralkyl, or heterocycloalkyl.

The term "sulfonate" refers to -SO2 _- OR, where R is -H, alkyl, aryl, aralkyl, or heterocycloalkyl.

The term "alkenyl" refers to an unsaturated group having 2 to 12 atoms and containing at least one carbon-carbon double bond, and includes straight chain, branched chain and cyclic groups. Alkenyl groups may be optionally substituted. Suitable alkenyl groups include allyl. "1-alkenyl" refers to an alkenyl group in which the double bond is between the first and second carbon atoms. If a 1-alkenyl group is attached to another group, for example, it is a W substituent attached to a cyclic phosphonate, it is attached at the first carbon.

The term "alkynyl" refers to an unsaturated group having 2 to 12 atoms and containing at least one carbon-carbon triple bond, and includes straight chain, branched chain and cyclic groups. Alkynyl groups may be optionally substituted. Suitable alkynyl groups include ethynyl. "1-alkynyl" refers to an alkynyl group in which the triple bond is between the first and second carbon atoms. If a 1-alkynyl group is attached to another group, for example, it is a W substituent attached to a cyclic phosphonate, it is attached at the first carbon.

The term "alkylene" refers to a divalent straight chain, branched chain or cyclic saturated aliphatic group. In one aspect, the alkylene group contains up to and includes 10 atoms. In another aspect, the alkylene group contains up to and includes 6 atoms. In another aspect, the alkylene group contains up to and includes 4 atoms. The alkylene group can be straight chain, branched chain or cyclic.

The term "acyloxy" refers to an ester group -O-C(O)R, where R is H, alkyl, alkenyl, alkynyl, aryl, aralkyl, or heterocycloalkyl.

The term "aminoalkyl" refers to the group _NR2 -alk-, wherein "alk" is an alkylene group and R is selected from -H, alkyl, aryl, aralkyl and heterocycloalkyl.

The term "alkylaminoalkyl" refers to alkyl-NR-alk-, wherein each "alk" is an independently selected alkylene group, and R is H or lower alkyl. "Lower alkylaminoalkyl" refers to a group in which the alkyl and alkylene groups are lower alkyl and alkylene groups, respectively.

The term "arylaminoalkyl" refers to the group aryl-NR-alk-, wherein "alk" is alkylene, and R is -H, alkyl, aryl, aralkyl or heterocycloalkyl. In "lower arylaminoalkyl", alkylene is lower alkylene.

The term "alkylaminoaryl-" refers to alkyl-NR-aryl-, wherein "aryl" is a divalent group and R is -H, alkyl, aralkyl or heterocycloalkyl. In "lower alkylaminoaryl", alkyl is lower alkyl.

The term "alkoxyaryl" refers to an aryl group substituted by an alkoxy group. In "lower alkoxyaryl", the alkyl group is a lower alkyl group.

The term "aryloxyalkyl" refers to an alkyl group substituted with an aryloxy group.

The term "aralkyloxyalkyl" refers to an aryl-alk-O-alk- group, wherein "alk" is alkylene. "Lower aralkyloxyalkyl" refers to a group wherein alkylene is lower alkylene.

The term "alkoxy-" refers to an alkyl-O- group.

The term "alkoxyalkyl" refers to an aryl-alk-O-alk- group, wherein "alk" is an alkylene group. In "lower alkoxyalkyl", each of alkyl and alkylene is a lower alkyl and alkylene group, respectively.

The term "alkylthio-" refers to the group alkyl-S-.

The term "alkylthioalkyl" refers to alkyl-S-alk-, wherein "alk" is alkylene. In "lower alkylthioalkyl", each of alkyl and alkylene is lower alkyl and alkylene, respectively.

The term "alkoxycarbonyloxy-" refers to alkyl-O-C(O)-O-.

The term "aryloxycarbonyloxy-" refers to aryl-O-C(O)-O-.

The term "alkylthiocarbonyloxy" refers to alkyl-S-C(O)-O-.

The term "amido" refers to an NR 2 group next to an acyl or sulfonyl group such as in NR ₂ -C(O)-, RC(O)-NR ¹ -, NR ₂ -S(=O) ₂ -, and RS(=O) ₂ -NR ¹ -, wherein R and R ¹ _include -H, alkyl, aryl, aralkyl, and heterocycloalkyl.

The term "amido" refers to NR ₂ -C(O)- and RC(O)-NR ¹ -, wherein R and R ¹ include -H, alkyl, aryl, aralkyl, and heterocycloalkyl. The term does not include urea-NR-C(O)-NR-.

The term "sulfonylamino" refers to _NR2 -S(=O) _2- and RS(=O) ₂ - ^NR1- , wherein R and ^R1 include -H, alkyl, aryl, aralkyl and heterocycloalkyl. The term does not include sulfonylurea-NR-S(=O) _2- NR-.

The terms "amidoalkylaryl" or "amidoaryl" refer to aryl-alk-NR ¹ -C(O) and ar-NR ¹ -C(O)-alk-, respectively, wherein "ar" is aryl, "alk" is alkylene, and R ¹ and R include H, alkyl, aryl, aralkyl, and heterocycloalkyl.

The term "sulfonamidoalkylaryl" or "sulfonamidoaryl" refers to aryl-alkyl-NR ¹ -S(=O) ₂ - and ar-NR ¹ -S(=O) ₂ -, respectively, wherein "ar" is aryl, "alk" is alkylene, and R ¹ and R 2 include -H, alkyl, aryl, aralkyl and heterocycloalkyl.

The term "hydroxyalkyl" refers to an alkyl group substituted with one -OH group.

The term "haloalkyl" refers to an alkyl group substituted with a halogen.

The term "cyano" refers to -CN.

The term "nitro" refers to -NO ₂ .

The term "acylalkyl" refers to alkyl-C(O)-alk-, where "alk" is alkylene.

The term "aminoamidoalkyl" refers to the group NR2 _- C(O)-N(R)-alk-, wherein R is alkyl or H, and "alk" is alkylene. "Lower aminoamidoalkyl" refers to the group wherein "alk" is lower alkylene.

The term "heteroarylalkyl" refers to an alkylene group substituted with a heteroaryl group. II. Compounds that interfere with PD-L1 activity

One aspect of the present application relates to compounds that interfere with PD-L1 activity. In certain embodiments, the compound has a general structure as shown in formula (I): (I)

or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate or tautomer thereof, wherein:

A and B are each independently selected from halogen, cyano, -N ₃ , alkyl and substituted alkyl, amine, alkylamine, alkoxy;

Z ₁ is -CR ¹ = or -N =;

Z ₂ is -CR ² =;

Z ₃ is -CR ³ = or -N =;

Z ₄ is -CR ⁴ = or -N =;

Z ₅ is -CR ⁵ =;

Z ₆ is -CR ⁶ = or -N =;

^R1 and ^R4 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

^R2 and ^R5 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

^R3 and ^R6 are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

_Y1 and _Y2 are independently -C( ^R7 )( ^R8 )-, ^-CR9 =, ^-NR10- , -O- or -S-;

_X1 and _X2 are each independently -C( ^R11 )( ^R12 )-, -N=, ^-NR13- , -S- or -O-;

R ⁷ , R ⁸ , R ⁹ , R ¹¹ and R ¹² are each independently -H, halogen, cyano, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, amine, alkylamine, alkoxy;

R ¹⁰ and R ¹³ are each independently -H, alkyl, cycloalkyl, substituted alkyl, alkenyl, alkynyl, aryl, alkylamine, alkoxy;

_L1 and _L2 are each an alkyl group, a substituted alkyl group or a heteroatom chain containing m atoms between ring 3 and _W1 and between ring 6 and _W2 , wherein m=0, 1, 2, 3, 4, 5 or 6; when m is 0, _W1 or _W2 is directly connected to the corresponding nitrogen in ring 3 or ring 6, respectively;

_W1 and _W2 are each independently hydrogen, a five-membered heterocyclic ring or a substituted five-membered heterocyclic ring, a six-membered heterocyclic ring or a substituted six-membered heterocyclic ring, a carboxyalkyl group or a substituted carboxyalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an amino acid ester, an amino acid amide, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid.

The compound of formula (I) may be symmetric about axis DD (i.e., the left part of formula (I) is a mirror image of the right part of formula (I)) or asymmetric (i.e., the left part of formula (I) is different from the right part of formula (I)). Preferred core structure

In certain embodiments, the compound of formula (I) comprises a core structure selected from formula (II)-(XXIII): (II) (III) (IV) (V) (VI) (VII) (VIII) (IX) (X) (XI) (XII) (XIII) (XIV) (XV) (XVI) (XVII) (XVIII) (XIX) (XX) (XXI) (XXII) (XXIII).

In certain embodiments, the compound of formula (I) comprises the following core structure:

and .

In certain embodiments, L ₁ and L ₂ are each independently selected from -CH ₂ -, (CH ₂ ) ₂ -, and -(CH ₂ ) ₃ -.

In certain embodiments, _L1 and _L2 are each independently selected from -CH( _CH3 )-, _-CH2- , and _-CH2 -CH( _CH3 )-. Type I Side Chain

In certain embodiments, _W1 and/or _W2 are each independently a type I side chain. As used herein, the term "type I side chain" refers to a structure containing: (1) a five-membered heterocyclic ring having at least one nitrogen atom as a ring atom or a substituted five-membered heterocyclic ring having at least one nitrogen atom as a ring atom, or (2) a six-membered heterocyclic ring having at least one nitrogen atom as a ring atom or a substituted five-membered heterocyclic ring having at least one nitrogen atom as a ring atom, wherein the linking group _L1 or _L2 is directly connected to a ring atom in the five-membered or six-membered heterocyclic ring.

In certain embodiments, W ₁ and/or W ₂ are each independently a heterocyclic ring. In certain embodiments, W ₁ and/or W ₂ are each independently a five-membered heterocyclic ring having at least one nitrogen atom as a ring atom. In certain embodiments, W ₁ and/or W ₂ are each independently a six-membered heterocyclic ring having at least one nitrogen atom as a ring atom.

In certain embodiments, _W1 and/or _W2 are each independently selected from the following I-type side chains:

, , , , ,

, , , , , , , , , , , , , , , , , , , , , , , and . Type II Side Chain

In certain embodiments, _W1 and/or _W2 are each independently a type II side chain. As used herein, the term "type II side chain" refers to a side chain having the general formula The _W1 structure and the general formula The _W2 structure,

wherein R ¹⁴ and R ¹⁶ are each independently -H, alkyl, substituted alkyl, hydroxyalkyl or substituted hydroxyalkyl, hydroxycarboxylic acid or a salt or ester thereof, substituted hydroxycarboxylic acid or a salt or ester thereof, carboxylic acid or a salt or ester thereof or an alkyl ether, substituted carboxylic acid or a salt or ester thereof or an alkyl ether, carbamide or lactone; and

wherein R ¹⁵ and R ¹⁷ are each independently -H, alkyl or substituted alkyl.

In certain embodiments, R ¹⁴ or R ¹⁶ or both are of the formula L ₃ -C(O)-Q ₂ R ¹⁸ , wherein L ₃ is alkyl, substituted alkyl, alkylamino or alkyl-amino-alkyl, Q ₂ is -O- or -CH ₂ -, and R ¹⁸ is -H, alkyl or substituted alkyl.

In certain embodiments, R ¹⁴ or R ¹⁶ or both are independently selected from:

, , , , , , , , , , , , , , , , , and .

In certain embodiments, _W1 or _W2 are each independently an amino acid.

In certain embodiments, side chain _W1 or _W2 is L-serine.

In certain embodiments, side chain _W1 or _W2 is L-serine.

In certain embodiments, side chains _W1 and _W2 are both L-serine.

In certain embodiments, side chains _W1 and _W2 are both L-serine.

In certain embodiments , _W1 or _{W2 are} each independently -C(O)-ONa, -CN, _-CH2OH or _-CH2NH2 .

In certain embodiments, the compound of formula (I) comprises a core structure:

.

In certain embodiments, the compound of formula (I) comprises a core structure:

, .

In certain embodiments, the compound of formula (I) is composed of two identical core structures linked together. composition.

In certain embodiments, the compound of formula (I) is composed of two identical core structures linked together. composition.

In certain embodiments, the compound of formula (I) is composed of a core structure connected together and core structure composition.

In certain embodiments, side chains _W1 and/or _W2 are independently selected from:

, , , , , and .

In certain embodiments, the compound of formula (I) comprises only one side chain, wherein:

W ₁ is , or ,

W ₂ is H, and L ₂ is absent (ie, m=0). In other words, there is no side chain at the nitrogen atom in ring 6. In another embodiment, L ₁ is C ₁ -C ₃ alkyl.

In certain embodiments, compounds of Formula (I) include side chains that are asymmetric about axis DD.

In certain embodiments, _W1 is , and W ₂ is selected from Type I and Type II side chains. In another embodiment, L ₁ and L ₂ are each independently C ₁ -C ₃ alkyl.

In certain embodiments, _W1 is , and W ₂ is selected from Type I and Type II side chains. In another embodiment, L ₁ and L ₂ are each independently C ₁ -C ₃ alkyl.

In certain embodiments, _W1 is , and W ₂ is selected from Type I and Type II side chains. In another embodiment, L ₁ and L ₂ are each independently C ₁ -C ₃ alkyl.

In certain embodiments, _W1 is , and _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, _W1 is , and _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, _W1 is , and _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

In certain embodiments, the side chain _W1 is , and the side chain _W2 is In another embodiment, _L1 and _L2 are each independently _C1 - _C3 alkyl.

Certain embodiments of the compounds disclosed herein are listed, without limitation, in Table 1 (or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, solvate or tautomer thereof). Table 1 Compound No. Structure [M+H] ⁺ 281 707.43 303 679.45 448 711.47 338 711.50 354 741.50 361 705.52 363 709.44 372 767.40 376 779.41 381 785.12 382 769.55 388 747.10 395 747.11 396 747.15 401 713.16 402 737.52 457 713.48 466 715.15 481 771.50 445 691.2 449 797.3 454 831.45 460 825.3 461 773.55 465 779.25 468 747.2 469 799.2 470 807.25 471 719.52 475 807.41 476 743.2 478 831.2 482 831.50 483 779.25 484 831.45 490 803.51 491 745.2 493 745.2 513 827.50 527 771.48 528 807.45 536 747.41 537 835.51 541 807.49 545 807.21 551 807.21 552 807.21 553 779.40 560 807.21 566 863.77 570 833.20 571 891.31 573 827.18 588 807.21 479 747.46 480 775.50 563 807.43 550 807.43 554 815.33 589 871.50 561 815.16 557 815.16 586 795.35 585 739.50 514 791.58 373 711.47 374 707.43 375 705.42 378 605.50 384 741.50 386 633.37 427 777.45 394 739.50 397 595.40 408 767.56 411 683.2 503 683.2 530 755.2 531 683.2 535 711.2 539 735.2 540 735.2 412 681.41 413 715.42 415 677.43 459 881.4 422 997.4 430 749.44 472 715.2 567 767.2 529 683.49 558 711.43 559 715.39 538 603.15 429 614.41 385 629.44 533 731.21 516 728.23 515 713.22 542 648.12 488 771.19 489 742.21 492 727.20 450 711.51 517 697.47

The present application also includes isotopically substituted compounds of the present disclosure. An "isotopically substituted" compound is a compound of the present application in which one or more atoms are replaced or substituted by atoms having the same atomic number but a different atomic mass or mass number, e.g., an atomic mass or mass number different from the atomic mass or mass number normally found in nature (i.e., naturally occurring). It should be understood that a "radiolabeled" compound is a compound into which at least one radioactive isotope (e.g., a radionuclide) has been incorporated. III. Uses of compounds of formula (I)

Another aspect of the present application relates to the use of the compound of formula (I). The compound of formula (I) interferes with the interaction between PD-L1 and PD-1, and thus can be used to treat diseases and disorders related to the activity of PD-1 and diseases and disorders related to PD-L1.

In certain embodiments, the compound of formula (I) promotes the formation of PD-L1 dimers and thereby inhibits the interaction between PD-L1 and PD-1. In certain embodiments, the compounds of the present disclosure or their pharmaceutically acceptable salts or stereoisomers are used for therapeutic administration to enhance, stimulate and/or increase immunity to cancer, chronic infection or sepsis, including enhancing the response to vaccination. In certain embodiments, the present disclosure provides a method for inhibiting PD-1/PD-L1 protein/protein interactions. The method comprises administering an effective amount of a compound of formula (I) or its pharmaceutically acceptable salt or stereoisomer to an individual or patient. The compound of formula (I) can be used alone, in combination with other agents or therapies, or as an adjuvant or novel adjuvant for the treatment of diseases or conditions including cancer or infectious diseases. For the uses described herein, any compound of the disclosure, including any embodiments thereof, may be used.

The compounds of the present application inhibit PD-1/PD-L1 protein/protein interactions, resulting in PD-1 pathway blockade. Blockade of PD-1 can enhance immune responses to cancer cells and infectious diseases in mammals, including humans. In certain embodiments, the present disclosure provides in vivo treatment of an individual or patient using a compound of any formula herein or a salt or stereoisomer thereof, so as to inhibit the growth of a cancerous tumor. A compound of any formula as described herein, a compound recorded in any claim and described herein, or a salt or stereoisomer thereof can be used to inhibit the growth of a cancerous tumor. Alternatively, a compound of any formula as described herein or a compound recorded in any claim and described herein, or a salt or stereoisomer thereof can be used in combination with other agents or standard cancer treatments as described below. In one embodiment, the disclosure provides a method for inhibiting the growth of tumor cells in vitro. The method comprises contacting tumor cells with a compound of any formula as described herein, or a compound or a salt or stereoisomer thereof as recorded in any claim and described herein in vitro. In another embodiment, the disclosure provides a method for inhibiting the growth of tumor cells in an individual or patient. The method comprises administering a therapeutically effective amount of a compound of any formula as described herein, or a compound or a salt or stereoisomer thereof as recorded in any claim and described herein to an individual or patient in need of the method.

In certain embodiments, provided herein is a method for treating cancer. The method comprises administering to a patient in need thereof a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof. Examples of cancer include cancers whose growth can be inhibited using the compounds disclosed herein and cancers that are generally responsive to immunotherapy.

Examples of cancers that can be treated using the compounds of the present disclosure include, but are not limited to: bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, perianal cancer, gastric cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, including acute Myeloid leukemia, chronic myeloid leukemia, acute lymphocytic leukemia, chronic or acute leukemia of chronic lymphocytic leukemia, solid tumors in children, lymphocytic lymphoma, bladder cancer, kidney or urethral cancer, renal pelvic cancer, central nervous system (CNS) tumor, primary CNS lymphoma, tumor angiogenesis, spinal cord tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermal cancer, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers including cancers induced by asbestos, and combinations of said cancers. The compounds disclosed herein can also be used to treat metastatic cancers, especially metastatic cancers expressing PD-L1.

In certain embodiments, cancers that can be treated using the compounds of the present disclosure include melanoma (e.g., metastatic malignant melanoma, cutaneous melanoma), kidney cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), breast cancer (e.g., invasive breast cancer), colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), squamous cell head and neck cancer (e.g., squamous cell carcinoma of the head and neck), urothelial cancer (e.g., bladder cancer, non-muscle invasive bladder cancer (NMIBC)), and cancers with high microsatellite instability (MSIhlgh).

Additionally, the present disclosure includes refractory or recurrent malignant tumors whose growth can be inhibited using the compounds of the present disclosure.

In certain embodiments, cancers that can be treated using the compounds of the present disclosure include, but are not limited to: solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), blood cancers (e.g., lymphoma, leukemia, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), DLBCL, mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma or multiple myeloma, and combinations of the aforementioned cancers.

In certain embodiments, cancers that can be treated using the compounds disclosed herein include, but are not limited to, cholangiocarcinoma, cholangiocarcinoma, biliary carcinoma, triple-negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing's sarcoma, brain cancer, brain tumor, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epithelioid Sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, islet cell cancer, oral cancer, oral cancer, mouth cancer, pharyngeal cancer, laryngeal cancer, lip cancer, mesothelioma, neck cancer, nasal cancer, eye cancer, ocular melanoma, pelvic cancer, rectal cancer, kidney cell cancer, salivary gland cancer, nasal sinus cancer, spinal cord cancer, tongue cancer, tubule cancer, urethra cancer and ureter cancer.

In certain embodiments, diseases and indications that can be treated using the compounds of the present disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancers, gastrointestinal cancers, genitourinary tract cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

Exemplary hematological cancers include lymphomas and leukemias, such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV) and essential thrombocythemia (ET)), myelodysplastic syndrome (MDS), T-cell acute lymphoblastic lymphoma (T-ALL) and multiple myeloma (MM).

Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyosarcoma, rhabdomyosarcoma, fibroma, lipoma, hamartoma, and teratoma.

Exemplary lung cancers include non-small cell lung cancer (NSCLC) (e.g., squamous cell NSCLC), small cell lung cancer, bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchogenic) carcinoma, bronchial adenoma, chondroitinoma, and mesothelioma.

Exemplary gastrointestinal cancers include esophageal cancer (carcinoma, squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma, adenocarcinoma), pancreatic cancer (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vasodilatory peptide tumor), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colorectal cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) and colorectal cancer (e.g., colorectal adenocarcinoma).

Exemplary genitourinary tract cancers include renal cancer (adenocarcinoma, Wilm's tumor [nephroblastoma]), bladder and urethral cancer (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate cancer (adenocarcinoma, sarcoma), and testicular cancer (seminoma, teratoma, embryonal carcinoma, teratoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma). In certain embodiments, the cancer is a urinary system tumor (e.g., papillary renal carcinoma, testicular germ cell carcinoma, renal chromophobe cell carcinoma, renal clear cell carcinoma, or prostate adenocarcinoma).

Exemplary liver cancers include hepatocellular carcinoma (liver cell carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

Exemplary bone cancers include, e.g., osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrohistocytic tumor, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondroma (osteocartilaginous exostosis), benign enchondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor.

Exemplary cancers of the nervous system include cranial cancer (osteomas, hemangiomas, granulomas, xanthomas, osteitis deformans), meningeal cancer (meningioma, meningiosarcoma, neuroglioma), brain cancer (astrocytoma, medulloblastoma, neuroglioma, ependymoma, germ cell tumor (pinealoma), glioblastoma, multiform glioblastoma, oligodendroglioma, neurothecoma, retinoblastoma, congenital tumors), and spinal cord cancer (neurofibroma, meningioma, neuroglioma, sarcoma), as well as neuroblastoma and Lhermitte-Duclos disease.

Exemplary gynecological cancers include uterine cancer (endometrial cancer), cervical cancer (cervical cancer, precancerous cervical atypical hyperplasia), ovarian cancer (ovarian cancer (serous cystadenocarcinoma, serous adenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), ovarian granulocytoma, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulvar cancer (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vaginal cancer (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), and fallopian tube cancer.

Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma (e.g., squamous cell carcinoma of the skin), Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermal fibroma, and keloid. In certain embodiments, diseases and indications that can be treated using the disclosed compounds include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, bile duct cancer, esophageal cancer, and urothelial carcinoma.

PD-1 pathway blockade using the disclosed compounds can also be used to treat infections, such as viral, bacterial, fungal and parasitic infections. In certain embodiments, methods for treating infections are provided herein. The method comprises administering to a patient in need of the method a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof. Examples of viruses that can be treated by the disclosed methods include, but are not limited to, human immunodeficiency virus, human papillomavirus, influenza, hepatitis A, B, C or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, coronavirus, severe acute respiratory syndrome virus, Ebola virus and measles virus. In certain embodiments, the methods disclosed herein can treat infections caused by viruses including, but not limited to, hepatitis (A, B, or C), herpes viruses (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein Barr virus), adenovirus, influenza virus, arbovirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum virus, poliovirus, rabies virus, JC virus, tuberculosis, and arbovirus encephalitis virus.

The present disclosure provides a method for treating bacterial infection, which comprises administering a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof to a patient in need of the method. Non-limiting examples of pathogenic bacteria that may be treated by the disclosed methods include: Chlamydia, Rickettsial , Mycobacteria , Staphylococci , Streptococci , Pneumonococci, Meningococci and Conococci , Klebsiella , Proteus , Serratia , Pseudomonas , Legionella , Diphtheria, Salmonella , Bacilli , and ), cholera, tetanus, botulism, anthrax, plague, leptospirosis, and Lyme disease bacteria.

The present disclosure provides a method for treating fungal infection, which comprises administering a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof to a patient in need of the method. Non-limiting examples of pathogenic fungi causing infections that can be treated by the disclosed methods include Candida ( albicans , krusei , glabrata , tropicalis, etc.), Cryptococcus neoformans , Aspergillus ( fumigatus , niger , etc.), Genus Mucorales ( mucor , absidia , rhizophus ), Sporothrix schenckii , Blastomyces dermatitidis , Paracoccidioides brasiliensis ), Coccidioides immitis and Flistoplasma capsulatum.

The present disclosure provides a method for treating parasitic infections, which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof. Non-limiting examples of pathogenic parasites that can be treated by the disclosed methods include: Entamoeba histolytica , Balantidium coli , Naegleria fowleri , Acanthamoeba sp. , Giardia lambia , Cryptosporidium sp ., Pneumocystis carinii , Plasmodium vivax , Babesia microti , Trypanosoma brucei , Trypanosoma cruzi , Leishmania donovani , Toxoplasma gondii, and gondi ) and Nippostrongylus brasiliensis .

In certain embodiments, provided herein is a method for treating inflammation, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof.

In certain embodiments, provided herein is a method for treating an autoimmune disease, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof.

The compounds of formula (I) or any of its embodiments may have desirable pharmacological properties and promising biomedical properties, such as toxicological properties, metabolic and pharmacokinetic properties, solubility and permeability. It should be understood that the determination of appropriate biomedical properties is within the knowledge of ordinary skill in the art, such as determining cell cytotoxicity or inhibiting certain targets or channels to determine potential toxicity.

In certain embodiments, the compounds of the present application can be used to prevent or reduce the risk of progression of any of the diseases referred to herein; for example, to prevent or reduce the risk of progression of a disease, disease state or disorder in an individual who may be susceptible to the disease, disease state or disorder but has not yet experienced or displayed the pathology or symptoms of the disease.

In certain embodiments, the disclosure provides a method for enhancing, stimulating and/or increasing an immune response in a patient. The method comprises administering to a patient in need thereof a therapeutically effective amount of a compound of any formula as described herein, a compound or composition described in any claim and described herein, or a salt thereof. Combination Therapy

The compounds disclosed herein can be used in combination with one or more therapies for treating a disease (e.g., cancer or infection). Examples of diseases and indications that can be treated using combination therapy include those described herein.

Examples of cancer include solid tumors and non-solid tumors, such as liquid tumors and blood cancers. Examples of infection include viral infection, bacterial infection, fungal infection or parasitic infection. For example, the compounds of the present disclosure can be combined with one or more inhibitors of the following kinases to treat cancer: Akt1, Akt2, Akt3, BCL2, CDK, TGF-PR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFotR, PDG1'PR, PI3K (α, β, γ, δ, and multiple or selective), CSF1R, KIT, FLK-1I, KDR/FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, PARP, Ron, Sea, I RKA, TRKB, TRKC, TAM kinases (Axl, Mer, Tyro3), FLT3, VEGFR/FH2, Flt4, EphAl, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf. In certain embodiments, the compounds of the present disclosure can be combined with one or more of the following inhibitors to treat cancer or infection. Non-limiting examples of inhibitors that can be combined with the compounds of the present disclosure to treat cancer and infection include FGFR inhibitors (FGFR1, FGFR2, FGFR3 or FGFR4, such as pemitinib (INCY54828), INCB62079), JAK inhibitors (JAK1 and/or JAK2, such as ruxolitinib, baricitinib or itacitinib (INCB39110)), IDO inhibitors (e.g., icadolstat, NLG919 or BMS-986205, MK7162), LSD1 inhibitors (e.g., INCB59872 and INCB60003), TDO inhibitors, PI3K-δ inhibitors (e.g., Parsaclisib (INCB50465) and INCB50797), PI3K-γ inhibitors (e.g., PI3K-γ selective inhibitors), Pirn inhibitors (e.g., INCB53914), EGFR inhibitors (also known as ErB-1 or HER-1; e.g., erlotinib, gefitinib, vandetanib, osimertinib, cetuximab, necituzumab, or panitumumab), VEGFR inhibitors or pathway blockers ( For example, bevacizumab, pazopanib, sunitinib, cetuximab, brucei, latinib), PARP inhibitors (e.g., olaparib, prinanib, venetinib, laclazimab, talazoparib, or niraparib), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl, and Mer), adenosine receptor antagonists (e.g., A2a/A2b receptor antagonists), HPK1 inhibitors, kinase receptor inhibitors (e.g., CCR2 or CCR5 inhibitors), SHP1/2 phosphatase inhibitors, histone deacetylase inhibitors (HD AC) (e.g., HDAC8 inhibitors), angiogenesis inhibitors, interleukin receptor inhibitors, bromodomain and extra terminal family member inhibitors (e.g., bromodomain inhibitors or BET inhibitors, such as INCB54329 and INCB57643), arginase inhibitors (INCB001158), PARP inhibitors (such as rucaparib or olaparib), sitravatinib, B-Raf inhibitor-MEK inhibitor combinations (e.g., encofenib plus bimetinib, dabrafenib plus trametinib or cobimetinib plus vemurafenib), and adenosine receptor antagonists or combinations thereof.

In certain embodiments, the compounds of the present disclosure can be combined with a TLR7 agonist (e.g., imiquimod).

The compounds disclosed herein can also be used in combination with other methods for treating cancer, such as chemotherapy, radiotherapy, tumor-targeted therapy, adjuvant therapy, immunotherapy or surgery. Examples of immunotherapy include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, single copy antibodies, bispecific or multispecific antibodies, antibody drug conjugates, T cell infusion, Toll receptor agonists, STING agonists, RIG-I agonists, oncolytic virus therapy, and immunomodulatory small molecules including thalidomide or JAK1/2 inhibitors, PI3K6 inhibitors, etc. The compounds can be administered in combination with one or more anticancer drugs such as chemotherapeutics. Examples of chemotherapeutic drugs include any of the following: abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, hexamethylmelamine, anastrozole, arsenic trioxide, asparaginase, atazacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, intravenous busulfan, oral busulfan, captestosterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, Actinomycin D, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin, dexrazoxane, docetaxel, doxorubicin, drostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibezetinib Mozambique, idarubicin, isocyclophosphamide, imatinib mesylate, interferon α-2a, irinotecan, lapatinib, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, nitrogen mustard, megestrol acetate, melphalan, tropurinol, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, norolone phenylpropionate, nelarabine, nofitumomab, oxaliplatin, paclitaxel, pamidronate disodium, panitumumab, pegaspargase, pegylated Filgrastim, pemetrexed disodium, pentostatin, guapobroman, plicamycin, procarbazine, rasburicase, rituximab, ruxolitinib, sorafenib, streptozotocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uramustine, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronic acid.

Other anticancer agents include antibody therapeutics, such as trastuzumab (Herceptin), antibodies to co-stimulatory molecules such as CTLA-4 (e.g., ipilimumab), 4-1BB (e.g., urelumab, utomilumab), antibodies to PD-1 and PD-L1, or antibodies to cytokines (IL-10, TGF-b, etc.). Examples of antibodies to PD-1 and/or PD-L1 that can be combined with the compounds of the present disclosure to treat cancer or infections such as viral, bacterial, fungal, and parasitic infections include, but are not limited to: nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, and SHR-1210.

The compounds of the present disclosure can be used in combination with one or more immune checkpoint inhibitors for the treatment of diseases such as cancer or infection.

Exemplary immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CBL-B, CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3K δ, PI3K γ, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TIGIT, CD112R, VISTA, PD-1, PD-L1, and PD-L2. In certain embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In certain embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3 and VISTA. In certain embodiments, the compounds provided herein can be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors and TGFR β inhibitors.

In certain embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

In certain embodiments, the inhibitor of the immune checkpoint molecule is a PD-1 inhibitor, such as an anti-PD-1 single copy antibody. In certain embodiments, the anti-PD-1 single copy antibody is nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001 or AMP-224. In certain embodiments, the anti-PD-1 single copy antibody is nivolumab or pembrolizumab. In certain embodiments, the anti-PD1 antibody is pembrolizumab.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab or tremelimumab.

In certain embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In certain embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, or INCAGN2385.

In certain embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In certain embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453 or TSR-022.

In certain embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of GITR, such as an anti-GITR antibody. In certain embodiments, the anti-GITR antibody is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323 or MEDI1873.

In some embodiments, the inhibitor of the immune checkpoint molecule is an inhibitor of OX40, for example, an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562, MOXR-0916, PF-04518600, GSK3174998 or BMS-986178. In some embodiments, the OX40L fusion protein is MEDI6383.

The compounds of the present disclosure can also be used in combination with one or more anti-inflammatory agents, steroids, immunosuppressants, or therapeutic antibodies.

A compound of any formula as described herein, a compound described in any claim and described herein, or a salt thereof can be combined with another immunogenic agent, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides and carbohydrate molecules), cells, and cells transfected with genes encoding immunostimulatory cytokines. Non-limiting examples of tumor vaccines that can be used include peptides of melanoma antigens, such as gplOO, MAGE antigens, Trp-2, MARTI and/or tyrosinase, or tumor cells transfected to express the cytokine GM-CSF.

Compounds of any formula as described herein, compounds described in any claim and described herein, or salts thereof, can be used in combination with an immune program for treating cancer. In certain embodiments, tumor cells are transduced to express GM-CSF. In certain embodiments, tumor vaccines include proteins from viruses involved in human cancers, such as human papilloma virus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's sarcoma herpes virus (KHSV). In certain embodiments, compounds of the present disclosure can be used in combination with tumor-specific antigens, such as heat shock proteins isolated from tumor tissue itself. In certain embodiments, compounds of any formula as described herein, compounds described in any claim and described herein, or salts thereof, can be combined with dendritic cell immunization to initiate a potent anti-tumor response.

The compounds disclosed herein can be used in combination with bispecific macrocyclic peptides that target effector cells expressing Fc α or Fc γ receptors to tumor cells. The compounds disclosed herein can also be combined with macrocyclic peptides that activate host immune responses.

The compounds disclosed herein can be used in combination with bone marrow transplants for the treatment of a variety of tumors of hematopoietic origin.

The compounds of any formula as described herein, the compounds recited in any claim and described herein, or salts thereof can be used in combination with vaccines to stimulate immune responses to pathogens, toxins and self-antigens.

When more than one agent is administered to a patient, they can be administered simultaneously, separately, sequentially, or in combination (e.g., for more than two agents). Formulations, dosage forms, and routes of administration

When used as a drug, the compounds of the present disclosure can be administered in the form of a pharmaceutical composition. Therefore, the present disclosure provides a composition comprising a compound of any formula as described herein, a compound recorded in any claim and described herein, or a pharmaceutically acceptable salt thereof, or any embodiment thereof, and at least one pharmaceutically acceptable carrier or formulation. These compositions can be prepared in a manner well known in the pharmaceutical field and can be administered by a variety of routes, depending on whether it is a local or systemic treatment and the area to be treated. Administration can be topical (including transdermal, epidermal, ocular, and mucosal, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols including through a nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, arterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion; or such as intrathecal, intracranial or intraventricular administration.

Parenteral administration may be in the form of a single bolus, or may be, for example, by continuous infusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners, etc. may be necessary or desirable.

The present application also includes a pharmaceutical composition containing a compound disclosed herein or a pharmaceutically acceptable salt thereof as an active ingredient in combination with one or more pharmaceutically acceptable carriers or excipients. In certain embodiments, the composition is suitable for topical administration. In the process of preparing the composition of the present application, the active ingredient is usually mixed with an excipient, which is diluted or enclosed in a carrier in the form of a capsule, a pouch, paper or other container. When an excipient is used as a diluent, it can be a solid, semi-solid or liquid material that serves as a vehicle, carrier or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, tablets, capsules, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (either as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

In preparing the formulation, the active compound can be ground to provide the appropriate fineness before combining with the other ingredients. If the active compound is substantially insoluble, it can be ground to a fineness of less than 200 mesh. If the active compound is substantially water-soluble, the fineness can be adjusted by grinding to provide a substantially uniform distribution in the formulation, for example, about 40 mesh.

The compounds of the present application can be ground using known grinding processes such as wet milling to obtain fineness suitable for tablet forming and other types of preparations. Finely divided (nanoparticle) preparations of the compounds of the present application can be prepared by methods known in the art, see for example WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gum tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup and methylcellulose. Formulations can also include: lubricants, such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives, such as methyl- and propylhydroxy-benzoates; sweeteners; and flavor correctors. The compositions of the present application can be formulated by procedures known in the art so as to provide quick, sustained or delayed release of the active ingredient after administration to a patient.

In certain embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein or a pharmaceutically acceptable salt thereof. In certain embodiments, the silicified microcrystalline cellulose comprises about 98% by weight microcrystalline cellulose and about 2% by weight silicon dioxide.

In certain embodiments, the composition is a sustained-release composition comprising at least one compound described herein or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or excipient. In certain embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose and polyethylene oxide. In certain embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, microcrystalline cellulose, lactose monohydrate and hydroxypropyl methylcellulose. In certain embodiments, the composition comprises at least one compound described herein or a pharmaceutically acceptable salt thereof, microcrystalline cellulose, lactose monohydrate and polyethylene oxide. In certain embodiments, the composition further comprises magnesium stearate or silicon dioxide. In certain embodiments, microcrystalline cellulose is Avicel PH102™. In certain embodiments, lactose monohydrate is Fast-flo 316™. In certain embodiments, hydroxypropylmethylcellulose is hydroxypropylmethylcellulose 2208 K4M (e.g., methylcellulose K4 M Premier ^™ ) and/or hydroxypropylmethylcellulose 2208 K100LV (e.g., methylcellulose K00LV ^™ ). In certain embodiments, polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Poly ox WSR 1105 ^™ ).

In certain embodiments, the composition is prepared using wet granulation. In certain embodiments, the composition is prepared using dry granulation.

The composition can be formulated in a unit dosage form, each dosage containing about 5 to about 1000 mg (1 g), more typically about 100 mg to about 500 mg of the active ingredient. In certain embodiments, each dosage contains about 10 mg of the active ingredient. In certain embodiments, each dosage contains about 50 mg of the active ingredient. In certain embodiments, each dosage contains about 25 mg of the active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable as a single dosage for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in association with a suitable drug formulation.

The components used to formulate the pharmaceutical composition are of high purity and substantially free of potentially harmful contaminants (e.g., at least national food grade, usually at least analytical grade, more typically at least pharmaceutical grade). Particularly for human consumption, the composition is preferably manufactured or formulated under good manufacturing practice standards as defined in applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations may be sterile and/or substantially isotonic and/or fully comply with all good manufacturing practice regulations of the U.S. Food and Drug Administration.

The active compound can be effective within a wide dosage range and is generally administered in a therapeutically effective amount. However, it should be understood that the actual amount of compound administered will generally be determined by a physician, depending on relevant circumstances including the disease state to be treated, the route of administration selected, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, etc.

The therapeutic dose of the compound of the present application can vary, for example, according to the specific use for treatment, the mode of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present application in the pharmaceutical composition can vary according to many factors including dosage, chemical properties (such as hydrophobicity) and route of administration. For example, the compound of the present application can be provided in an aqueous physiological buffer solution containing about 0.1% to about 10% w/v of the compound for parenteral administration. Some typical dosage ranges are about 1 pg/kg body weight to about 1 g/kg body weight per day. In certain embodiments, the dosage range is about 0.01 mg/kg body weight to about 100 mg/kg body weight per day. The dosage may depend on such variables as the type and extent of progression of the disease or condition, the general health of the particular patient, the relative biological potency of the compound selected, the formulation of the dosage form and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

In order to prepare solid compositions such as tablets, the main active ingredient is mixed with a drug excipient to form a solid preformulated composition containing a uniform mixture of the compound of the present application. When these preformulated compositions are referred to as uniform, the active ingredient is usually evenly dispersed throughout the composition, so that the composition can be easily subdivided into unit dosage forms of the same effect, such as tablets, pills and capsules. The solid preformulation is then subdivided into unit dosage forms of the above type containing, for example, about 0.1 to about 1000 mg of the active ingredient of the present application.

The tablet or pill of the present application can be coated or otherwise mixed to provide a dosage form that gives a long-acting advantage. For example, the tablet or pill can include an inner dose component and an outer dose component, and the outer dose component is in the form of encapsulating the inner dose component. The two components can be separated by an enteric layer, which is used to resist decomposition in the stomach and allows the inner component to be completely delivered to the duodenum or delayed when released. Various materials can be used for such enteric layers or coatings, including a variety of polymeric acids and a mixture of polymeric acids with materials such as wormwood, cetyl alcohol and cellulose acetate.

Liquid dosage forms for oral or injection administration into which the compounds and compositions of the present application can be incorporated include aqueous solutions, appropriately flavored syrups, aqueous or oily suspensions and flavored emulsions containing edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical carriers.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable formulations as described above. In certain embodiments, the composition is administered by the oral or nasal respiratory route for local or systemic action. The composition can be atomized by the use of an inert gas. The atomized solution can be inhaled directly from the atomizing device, or the atomizing device can be connected to a mask, face tent or intermittent positive pressure ventilator. The solution, suspension or powder composition can be administered orally or nasally from a device for delivering the formulation in an appropriate manner.

Topical preparations can contain one or more conventional carriers. In certain embodiments, ointments can contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white vaseline, etc. The carrier composition of a cream can be based on water combined with glycerol and one or more other components such as glyceryl monostearate, PEG-glyceryl monostearate, and hexadecyl stearyl alcohol. Gels can be prepared using isopropyl alcohol and water, appropriately combined with other components such as glycerol, hydroxyethyl cellulose, etc. In certain embodiments, a topical preparation contains at least about 0.1% by weight, at least about 0.25% by weight, at least about 0.5% by weight, at least about 1% by weight, at least about 2% by weight, or at least about 5% by weight of the present application compound. The topical formulation can be suitably packaged in, for example, a 100 g tube, optionally in combination with instructions for use in the treatment of a selected indication such as psoriasis or other skin disease state.

The amount of the compound or composition administered to a patient will vary depending on the substance being administered, the purpose of the administration, such as prevention or treatment, the condition of the patient, the mode of administration, and the like. In therapeutic applications, the composition can be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease condition being treated and the judgment of the attending physician, which depends on factors such as the severity of the disease, the age, weight and general condition of the patient, and the like.

The composition administered to the patient can be in the form of the above-mentioned pharmaceutical composition. These compositions can be sterilized by conventional sterilization techniques, or can be sterile filtered. Aqueous solutions can be packaged for use or lyophilized, and lyophilized preparations are combined with sterile aqueous carriers before administration. The pH of the compound preparation is generally 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It should be understood that the use of some of the aforementioned excipients, carriers or stabilizers will lead to the formation of drug salts.

The therapeutic dose of the compound of the present application can vary according to, for example, the specific use for treatment, the method of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compound of the present application in the pharmaceutical composition can vary according to many factors including dosage, chemical properties (such as hydrophobicity), and route of administration. For example, the compound of the present application can be provided in an aqueous physiological buffer solution containing about 0.1% to about 10% w/v of the compound for parenteral administration. Some typical dosage ranges are from about 1 pg/kg body weight to about 1 g/kg body weight per day. In certain embodiments, the dosage range is from about 0.01 mg/kg body weight to about 100 mg/kg body weight per day. The dosage may depend on such variables as the type and extent of progression of the disease or condition, the general health of the particular patient, the relative biological potency of the compound selected, the formulation of the dosage form and its route of administration. Effective doses can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

The present application is further illustrated by the following examples, which should not be construed as limiting. The contents of all references, patents and published patent applications, as well as the figures and tables cited in this application are incorporated herein by reference. Examples Example 1 Preparation of (S)-(5-oxopyrrolidin-2-yl)methyl-4-methylbenzenesulfonate (Intermediate SM1)

A solution of (S)-5-(Hydroxymethyl)pyrrolidin-2-one (0.100 g, 0.869 mmol, 1 eq), TsCl (0.182 g, 0.954 mmol, 1.10 eq), TEA (0.132 g, 1.306 mmol, 1.5 eq) and DMAP (0.006 g, 0.049 mmol, 0.05 eq) in DCM (2 ml) was stirred at room temperature for 16 h, 1 N HCl (5 ml) was slowly added, and the mixture was extracted with DCM. The organics were concentrated to give SM1 (0.170 g, 73%). Example 2 Preparation of 2,2'-(2,2'- dichloro- [1,1'- biphenyl ]-3,3'- diyl ) bis (4,4,5,5 -tetramethyl -1,3,2- dioxaborolane ) ( Intermediate SM2) Steps 1 and 2

3-Bromo-2-chlorophenol (12.4 g, 0.060 mol, 1.0 eq), B ₂ Pin ₂ (16.4 g, 0.065 mol, 1.08 eq), KOAc (20.5 g, 0.210 mol, 3.5 eq), Pd(dppf)Cl ₂ ·DCM (4.1 g, 5.1 mmol, 0.085 eq) were added to a 500 ml round-bottom flask, and then solvent dioxane (300 ml) was added, and N ₂ was added to the final mixture and stirred at 95° C. for 3 hours. The reaction mixture was then cooled to room temperature, filtered, the filter cake was washed with dioxane (100 ml), and the filtrate was used directly in the next step.

3-Bromo-2-chlorophenol (12.0 g, 0.059 mol, 0.99 eq), K ₂ CO ₃ (24.8 g, 0.180 mol, 3.0 eq) and Pd(dppf)Cl ₂ ·DCM (2.1 g, 2.40 mmol, 0.042 eq) were added to the previous filtrate, and then H ₂ O (80 ml) was added, and N ₂ was added to the final mixture and stirred at 85° C. for 3.5 hours. The reaction mixture was then cooled to room temperature and filtered, and the filter cake was washed with EA (300 ml). Brine (300 ml) was added to the filtrate and separated, the aqueous phase was extracted with EA (100 ml x2), and the combined organic phases were decolorized with activated carbon at room temperature overnight. The mixture was filtered using a celite pad, the filter cake was washed with EA, and the combined organic phases were concentrated in vacuo. The residue was purified by recrystallization with DCM/PE = 1.5/1 to afford the desired product (10.1 g, yield: 46%) as a light yellow solid. Step 3

To a stirred mixture of SM2-02 (10.1 g, 0.039 mol, 1.0 eq) in DCM (200 ml) at 0°C was added DIPEA (19.4 g, 0.151 mol, 3.8 eq). After SM2-02 was dissolved, Tf ₂ O (26.8 g, 0.095 mol, 2.4 eq) was added at this temperature, and then stirred at room temperature for another 2 hours. Water (100 ml) was added to cause a reaction, and then separated, and the aqueous phase was extracted with DCM (100 ml). The combined organic phases were washed with brine (200 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by recrystallization with EtOH/H ₂ O=1/1 to obtain the desired product (18.3 g, yield 89%) as a light yellow solid. Step 4

To a stirred solution of SM2-03 (14.2 g, 0.027 mol, 1.0 eq) in dioxane (80 ml) were added B ₂ Pin ₂ (27.8 g, 0.109 mol, 4.0 eq), KOAc (16 g, 0.164 mol, 6.0 eq) and Pd(dppf)Cl ₂ ·DCM (3.3 g, 4.1 mmol, 0.15 eq) at room temperature, and then N ₂ was added and stirred at 85° C. for 2 hours. Then, the mixture was cooled to room temperature, EA (150 ml) and water (150 ml) were added to the mixture, separated, and the aqueous phase was extracted with EA (100 ml). The combined organic phases were washed with brine (200 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was dissolved with EA (50 ml), and then PE (300 ml) was slowly added to the solution to form a black suspension. After stirring for 30 minutes, the mixture was filtered, the filter cake was washed with 140 ml (PE/EA = 6/1), and the filtrate was concentrated in vacuo. The residue was recrystallized with EtOH (150 ml) to obtain the desired product (9.5 g, yield: 75%) as an off-white solid. Example 3 Preparation of 2,2'-((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazine-7,4-diyl))diacetaldehyde (Intermediate SM3) Step 1

A solution of 7-bromo-2H-benzo[b][1,4]oxazin-3(4H)-one (5.00 g, 21.93 mmol, 1 eq), bromoacetaldehyde dimethyl acetal (5.55 g, 32.89 mmol, 1.5 eq) and Cs ₂ CO ₃ (14.29 g, 43.86 mmol, 2.0 eq) in DMF (60 ml) was stirred at 60°C for 16 hours, 180 ml of H ₂ O was added, and the mixture was extracted with EA. The organic layer was collected and purified with silica gel to give 7-bromo-4-(2,2-dimethoxyethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (5.40 g, 78%). Step 2

A solution of compound SM2 (2.50 g, 5.26 mmol, 1 eq), 7-bromo-4-(2,2-dimethoxyethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (3.49 g, 11.04 mmol, 2.1 eq), K ₂ CO ₃ (2.90 g, 21.04 mmol, 4.0 eq), Pd(dppf)Cl ₂ DCM (0.21 g, 0.263 mmol, 0.05 eq) in dioxane (40 ml) and H ₂ O (8 ml) was stirred at 80° C. under N ₂ for 2 h, and 30 mL of H ₂ O, extracted with EA, the organic layer was collected and purified with silica gel to obtain 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-(2,2-dimethoxyethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one) (3.10 g, 85%). Step 3

A solution of 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(4-(2,2-dimethoxyethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one) (3.10 g, 4.47 mmol, 1 eq) in 1 N HCl water/dioxane (15 ml/30 ml) was stirred at 80°C for 1 hour, 30 mL of _H2O was added, extracted with EA, the organic layer was washed with _NaHCO3 and concentrated to give the title compound SM3 (2.91 g, crude).

Preparation of 3,3'-((2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazine-7,4-diyl))dipropanal (intermediate SM4)

Compound SM4 can be prepared by following the same process as SM3 using 3-bromo-1,1-dimethoxypropane as a reactant. Example 4 Preparation of 7,7'-(2,2'- dichloro- [1,1' -biphenyl ]-3,3' -diyl ) bis (2H- benzo [b][1,4] oxazin -3(4H) -one ) ( Intermediate SM5)

Referring to the following reaction equation, SM2 (0.05 g, 0.11 mmol, 1 eq), 7-bromo-2H-benzo[b][1,4]oxazin-3(4H)-one (0.062 g, 0.27 mmol, 2.5 eq), Pd(dppf)Cl ₂ (0.008 g, 0.01 mmol, 0.1 eq) and potassium carbonate (0.058 g, 0.42 mmol, 4 eq) were dissolved in dioxane/H ₂ O (3 ml, v/v = 5:1). The reaction was carried out at 85° C. for 2 hours under N ₂ atmosphere. After cooling, 10 ml of water and 10 ml of EA were added for extraction, and the organic phase was concentrated and purified using preparative TLC (elution with DCM/MeOH = 10/1) to obtain compound SM5, 7,7'-(2,2'-dichloro-[1,1'-biphenyl]-3,3'-diyl)bis(2H-benzo[b][1,4]oxazin-3(4H)-one) (13 mg, yield: 15.6%). Example 5 ( Preparation by reductive amination ) 5A. Preparation of 1-(2-(7-(2,2'- dichloro -3'-(4-(2-((S)-3- hydroxypyrrolidin- 1- yl ) ethyl )-3- oxo -3,4 - dihydro - 2H- benzo [b][1,4] oxazin -7- yl )-[1,1'- biphenyl ]-3 - yl )-3- oxo -2,3- dihydro -4H- benzo [b][1,4] oxazin -4- yl ) ethyl ) pyrrolidine -3- carboxylic acid

To a stirred solution of compound SM3 (0.015 g, 0.024 mmol, 1 eq), pyrrolidine-3-carboxylic acid (0.0035 g, 0.03 mmol, 1.3 eq), (S)-pyrrolidin-3-ol hydrochloride (0.004 g, 0.032 mmol, 1.3 eq) and one drop of AcOH (1 mL/0.5 mL) in CH ₂ Cl ₂ /MeOH at room temperature was added sodium triacetoxyhydroborate (0.051 g, 0.24 mmol, 10 eq). After 4 hours, the mixture was directly concentrated and 0.5 mL of H ₂ O and 3 mL of MeOH were added, and the mixture was purified using reverse phase HPLC (0.1% trifluoroacetic acid in water/acetonitrile) to provide GLC01-481 (6 mg, 31%).

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (d, J = 7.8 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.19 (d, 2H), 7.14 (d, J = 2.0 Hz, 2H), 5.53 (s, 2H), 4.75 (s, 4H), 4.49 – 4.43 (m, 2H), 4.34 – 4.26 (m, 4H), 3.75 – 3.67 (m, 4H), 3.24 – 3.17 (m, 4H), 2.34 – 2.16 (m, 4H), 1.57 – 1.35 (m, 4H).

LCMS (ESI): for C ₄₁ H ₄₀ Cl ₂ N ₄ O ₇ ; [M+H] ⁺ : calcd: 771.23, found: 771.50 5B. The following compounds can be prepared using different amine substrates and SM3 or SM4 :

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.45 (d, J = 7.5 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.3 Hz, 2H), 7.14 (s, 2H), 4.75 (s, 4H), 4.29 (s, 4H), 3.66 (s, 2H) 3.35-3.29 (m, 8H), 2.29-2.20 (m, 8H).

LCMS (ESI): for C ₃₆ H ₃₆ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 691.2, found: 691.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.62 (s, 2H), 7.52-7.44 (m, 5H), 7.38 (dd, J = 17.4 Hz, 7.9 Hz, 4H), 7.16 (d, J = 8.4 Hz, 2H), 7.10 (s, 2H), 4.70 (s, 4H), 3.99 (td, J = 14.8 Hz, 7.5 Hz, 4H), 3.55 (dd, J = 12.1 Hz, 6.0 Hz, 2H), 2.76 (s, 4H), 2.55 (d, J = 5.7 Hz, 4H), 2.15-1.96 (m, 8H), 1.71-1.61 (m, 2H).

LCMS (ESI): for C ₄₂ H ₄₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 797.3, found: 797.3

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.6, 1.8 Hz, 2H), 7.40 (d, J = 7.3 Hz, 2H) , 7.35 (d, J = 8.4 Hz, 2H), 7.19 – 7.15 (m, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.36 (s, 2H), 4.31 – 4.14 (m, 6H), 3.72 – 3.64 (m, 4H), 3.12 – 2.97 (m, 4H), 2.20 – 2.08 (m, 4H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H]+: calcd: 831.22, found: 831.45

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.48 (dt, J = 13.7, 7.5 Hz, 6H), 7.40 (d, J = 7.4 Hz, 2H), 7.33 (d, J = 8.3 Hz, 2H) , 7.17 (d, J = 8.5 Hz, 2H), 7.11 (s, 2H), 4.69 (s, 4H), 4.08 (s, 2H), 4.02 (s, 2H), 3.50 (s, 2H), 2.63 ( s, 2H), 2.58 (d, J = 6.6 Hz, 4H), 2.27 (s, 6H), 2.14 – 1.92 (m, 8H), 1.65 – 1.57 (m, 2H).

LCMS (ESI): for C ₄₄ H ₄₆ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 825.3, found: 825.3

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.18 (t, J = 5.8 Hz, 2H), 7.52 (t, J = 7.6 Hz, 2H), 7.45 (d, J = 7.6 Hz, 2H), 7.41 (d, J = 7.3 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.17 (dd, J = 8.3, 1.9 Hz, 2H), 7.13 (s, 2H), 4.74 (s, 4H) , 4.24 (t, J = 6.7 Hz, 4H), 3.32 (q, J = 6.2 Hz, 5H), 3.23 (d, J = 8.0 Hz, 4H), 3.07 (t, J = 5.9 Hz, 4H), 1.84 (s, 6H).

LCMS (ESI): for C ₄₀ H ₄₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 773.26, found: 773.55

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.7, 1.9 Hz, 2H), 7.41 (dd, J = 7.4, 1.8 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.19 (dd, J = 8.4, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.38 – 4.18 (m, 4H), 4.04 (s, 2H), 3.97 – 3.77 (m, 4H), 3.22 (t, J = 7.1 Hz, 4H), 2.53 – 2.51 (m, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 779.19, found: 779.25

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.46 (d, J = 7.6 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.5 Hz, 2H), 7.13 (s, 2H), 4.74 (s, 4H), 4.24 (t, J = 5.8 Hz, 4H), 3.24 – 3.15 (m, 8H), 2.63 (t, J = 6.8 Hz, 4H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 747.2, found: 747.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.45 (d, J = 7.6 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.3 Hz, 2H), 7.14 (s, 2H), 4.75 (s, 4H), 4.30 (s, 4H), 3.50-3.30 (m, 14H), 2.20 (d, J = 63.2 Hz, 4H)

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 799.2, found: 799.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.46 (d, J = 7.4 Hz, 2H), 7.42 – 7.38 (m, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.2 Hz, 2H), 7.13 (d, J = 1.9 Hz, 2H), 4.74 (s, 4H), 4.34 – 4.18 (m, 4H), 4.15 ( s, 2H), 3.20 (d, J = 6.9 Hz, 4H), 3.13 (d, J = 12.2 Hz, 2H), 2.97 (t, J = 11.0 Hz, 2H), 2.45 (d, J = 5.4 Hz, 4H), 2.39 – 2.36 (m, 4H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.22, found: 807.25

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 9.74 (s, 2H), 7.52 (t, J = 7.6 Hz, 2H), 7.48 – 7.44 (m, 2H), 7.43 – 7.38 (m, 4H), 7.19 (dd, J = 8.4, 2.0 Hz, 2H), 7.14 (d, J = 2.0 Hz, 2H), 5.42 (s, 2H), 4.75 (s, 4H), 4.34 (t, J = 7.3 Hz, 4H ), 3.76 (t, J = 5.2 Hz, 4H), 3.50 - 3.40 (m, 4H), 3.33 - 3.16 (m, 4H), 2.93 (s, 6H).

LCMS (ESI): for C ₃₈ H ₄₀ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 719.23, found: 719.52

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51 (t, J = 7.6 Hz, 2H), 7.46 (dd, J = 7.6, 1.9 Hz, 2H), 7.40 (dd, J = 7.3, 1.9 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.19 (dd, J = 8.3, 2.0 Hz, 2H), 7.13 (d, J = 2.1 Hz, 2H), 4.72 (s, 4H), 4.25 ( s, 4H), 3.75 (d, J = 11.7 Hz, 2H), 3.60 (d, J = 11.6 Hz, 2H), 3.11 (d, J = 8.6 Hz, 4H), 2.65 – 2.55 (m, 4H), 1.35 (s, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.22, found: 807.41

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 – 7.44 (m, 4H), 7.39 (d, J = 7.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 7.17 (d, J = 8.3 Hz, 2H), 7.11 (s, 2H), 4.70 (s, 6H), 4.18 (s, 2H), 4.03 (t, J = 7.0 Hz, 4H), 2.80 (dd, J = 9.4, 6.2 Hz, 2H), 2.67 – 2.60 (m, 6H), 2.38 (dd, J = 9.9, 3.4 Hz, 2H), 2.03 – 1.91 (m, 4H), 1.53 (dd, J = 8.5, 4.2 Hz, 2H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 743.2, found: 743.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 – 7.44 (m, 4H), 7.39 (d, J = 7.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 7.17 (d, J = 8.3 Hz, 2H), 7.11 (s, 2H), 4.70 (s, 5H), 4.24 -4.18 (m, 3H), 4.03 (t, J = 7.0 Hz, 2H) 3.24 – 3.15 (m, 4H) , 2.80 (dd, J = 9.4, 6.2 Hz, 1H), 2.67-2.60 (m, 5H).2.38 (dd, J = 9.9, 3.4 Hz, 1H), 2.03 – 1.91 (m, 2H), 1.53 (dd, J = 8.5, 4.2 Hz, 1H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 831.2, found: 831.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.8, 1.9 Hz, 2H), 7.40 (dd, J = 7.5, 1.9 Hz, 2H), 7.33 (d, J = 8.5 Hz, 2H), 7.17 (dd, J = 8.2, 2.0 Hz, 2H), 7.12 (d, J = 1.9 Hz, 2H), 5.30 (s, 2H), 4.72 ( s, 4H), 4.33 – 4.29 (m, 2H), 4.20 – 4.12 (m, 4H), 4.07 – 3.87 (m, 4H), 3.59 – 3.56 (m, 2H), 3.07 – 3.02 (m, 2H), 2.88 – 2.82 (m, 2H), 2.10 – 2.03 (m, 4H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 831.21, found: 831.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.7, 1.9 Hz, 2H), 7.41 (dd, J = 7.4, 1.8 Hz, 2H), 7.37 (d, J = 8.5 Hz, 2H), 7.19 (dd, J = 8.4, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.38 – 4.18 (m, 4H), 4.04 (s, 2H), 3.97 – 3.77 (m, 4H), 3.22 (t, J = 7.1 Hz, 4H), 2.53 – 2.51 (m, 2H).

LCMS (ESI): for C ₃₆ H ₃₆ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 779.19, found: 779.25

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (d, J = 7.6 Hz, 2H), 7.41 (d, J = 7.7 Hz, 4H), 7.19 (d, J = 8.4 Hz, 2H), 7.13 (d, J = 2.1 Hz, 2H), 4.73 (s, 4H), 4.39 (s, 2H), 4.35 - 4.25 (m, 6H), 3.55 - 3.51( m, 4H), 3.25 – 3.20 (m, 4H), 2.65 – 2.60 (m, 2H), 2.07 (d, J = 13.5 Hz, 2H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 831.21, found: 831.45

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 12.16 (s, 2H), 8.56 (s, 4H), 7.52 (t, J = 7.6 Hz, 2H), 7.45 (d, J = 7.5 Hz, 2H) , 7.41 (d, J = 7.4 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.2, 2.0 Hz, 2H), 7.14 (d, J = 1.9 Hz, 2H) , 4.75 (s, 4H), 4.24 (t, J = 6.5 Hz, 4H), 3.23 – 3.16 (m, 4H), 3.03 – 2.97 (m, 4H), 2.27 (t, J = 7.0 Hz, 4H), 1.62 – 1.52 (m, 8H).

LCMS (ESI): for C ₄₂ H ₄₄ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 803.25, found: 803.51

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 – 7.44 (m, 4H), 7.39 (d, J = 7.3 Hz, 2H), 7.29 (d, J = 8.3 Hz, 2H), 7.17 (d, J = 8.3 Hz, 2H), 7.11 (s, 2H), 4.70 (s, 4H), 4.24 -4.18 (m, 3H), 4.03 (t, J = 7.0 Hz, 2H) 3.24 – 3.15 (m, 4H) , 2.80 (dd, J = 9.4, 6.2 Hz, 1H), 2.67-2.60 (m, 5H).2.38 (dd, J = 9.9, 3.4 Hz, 1H), 2.03 – 1.91 (m, 2H), 1.53 (dd, J = 8.5, 4.2 Hz, 1H).

LCMS (ESI): for C ₃₉ H ₃₈ Cl ₂ N ₄ O ₇ ; [M+H] ⁺ : calcd: 745.2, found: 745.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.5 Hz, 2H), 7.45 (d, J = 7.5 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.3 Hz, 2H), 7.14 (s, 2H), 4.75 (s, 4H), 4.29 (s, 4H), 3.65 (s, 1H) , 3.40-3.20 (m, 11H), 3.10 (s, 1H), 2.23 (s, 4H)

LCMS (ESI): for C ₃₉ H ₃₈ Cl ₂ N ₄ O ₇ ; [M+H] ⁺ : calcd: 745.2, found: 745.2

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.50 (t, J = 7.5 Hz, 2H), 7.47 – 7.44 (m, 2H), 7.41 (t, J = 7.9 Hz, 4H), 7.18 (dd, J = 8.4, 2.0 Hz, 2H), 7.12 – 7.09 (m, 2H), 4.67 (s, 4H), 4.08 (t, J = 6.4 Hz, 4H), 3.32 (t, J = 6.0 Hz, 2H), 3.01 – 2.93 (m, 4H), 2.92 – 2.81 (m, 4H), 2.74 – 2.66 (m, 2H), 1.91 – 1.81 (m, 4H), 1.67 – 1.48 (m, 6H).

LCMS (ESI): for C ₄₄ H ₄₄ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 827.25, found: 827.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.53-7.48 (m, 1H), 7.46 (dd, J = 7.6, 1.9 Hz, 1H), 7.40 (dd, J = 7.4, 1.9 Hz, 1H), 7.35 (d, J = 8.4 Hz, 1H), 7.16 (dd, J = 8.3, 2.0 Hz, 1H), 7.10 (d, J = 2.1 Hz, 1H), 4.70 (s, 2H), 4.29 (dd, J = 9.2, 5.4 Hz, 1H), 4.02 (dd, J = 9.3, 2.9 Hz, 1H), 3.98 (t, J = 7.2 Hz, 2H), 3.57 (s, 1H), 2.82-2.66 (m, 4H), 2.24 (dd, J = 17.2, 3.3 Hz, 1H).

LCMS (ESI): for C ₄₀ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 771.20, found: 771.48

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51-7.43 (m, 2H), 7.38 (dd, J = 7.8, 1.9 Hz, 1H), 7.31 (d, J = 8.3 Hz, 1H), 7.15 ( d, J = 8.3 Hz, 1H), 7.11 (d, J = 2.1 Hz, 1H), 4.82 (s, 1H), 4.70 (s, 2H), 4.01-3.94 (m, 2H), 3.59-3.44 (s , 3H), 2.83-2.64 (m, 4H), 2.25-2.20 (m, 1H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.22, found: 807.45

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52-7.46 (m, 1H), 7.45 (d, J = 7.6 Hz, 1H), 7.40 (dd, J = 7.4, 2.1 Hz, 1H), 7.35 ( d, J = 8.2 Hz, 1H), 7.16 (dd, J = 8.2, 2.0 Hz, 1H), 7.10 (d, J = 2.1 Hz, 1H), 4.69 (s, 2H), 3.98 (t, J = 7.2 Hz, 2H), 3.41-3.33 (m, 1H), 2.78-2.66 (m, 2H), 1.29 (d, J = 6.6 Hz, 3H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 747.20, found: 747.41

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.50 (t, J = 7.5 Hz, 2H), 7.48 - 7.43 (m, 2H), 7.39 (dd, J = 7.4, 1.7 Hz, 2H), 7.35 ( d, J = 8.5 Hz, 2H), 7.17 - 7.13 (m, 2H), 7.11 - 7.07 (m, 2H), 4.81 (t, J = 5.6 Hz, 2H), 4.69 (s, 4H), 4.07 (q , J = 7.1 Hz, 4H), 4.02 - 3.91 (m, 4H), 3.59 -3.49 (m 4H), 3.32 - 3.28 (m, 2H), 2.88 - 2.78 (m, 2H), 2.72 - 2.65 (m, 2H), 1.16 (t, J = 7.1, 1.0 Hz, 6H).

LCMS (ESI): for C ₄₂ H ₄₄ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 835.24, found: 835.51

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 9.36 (s, 3H), 7.52 (t, J = 7.5 Hz, 2H), 7.46 (d, J = 6.9 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 7.19 (d, J = 8.4 Hz, 2H), 7.13 (d, J = 1.9 Hz, 2H), 5.75 (s, 2H), 4.73 (s, 4H), 4.40 – 4.21 (m, 6H), 3.95 – 3.87 (m, 4H), 3.78 (s, 6H), 3.30 – 3.22 (m, 4H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.49

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.51-7.45 (m, 4H), 7.40-7.37 (m, 4H), 7.15 (d, J = 8.0 Hz, 2H), 7.10 (s, 2H), 4.70 (s, 4H), 4.16 (m, 2H), 4.08 (m, 2H), 3.85 (m, 2H), 3.08 (d, J = 4 Hz, 2H), 3.04 (m, 2H), 2.87 (m , 2H), 1.13 (d, J = 8.0 Hz, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.21

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.53-7.43 (m, 4H), 7.41-7.36 (m, 4H), 7.20 (d, J = 8.0 Hz, 2H), 7.12 (s, 2H), 4.71 (s, 4H), 4.15 (m, 2H), 4.09 (m, 2H), 3.87 (m, 2H), 3.10 (d, J = 4 Hz, 2H), 3.05 (m, 2H), 2.90 (m , 2H), 1.14 (d, J = 8.0 Hz, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.21

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.52-7.35 (m, 10H), 7.16 (d, J = 8.0 Hz, 2H), 7.11 (s, 2H), 7.02-6.82 (m, 2H), 4.74 (s, 4H), 4.10 (m, 4H), 3.51 (m, 4H), 2.99 (t, J = 6 Hz, 4H), 2.87 (m, 2H), 1.78 (m, 2H), 1.67 (m , 2H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.21

¹ H NMR (500 MHz, DMSO- d ₆ ) δ7.53 (t, J = 7.5 Hz, 2H), 7.48 (d, J = 6.9 Hz, 2H), 7.43 (d, J = 7.5 Hz, 2H), 7.38 (d, J = 8.4 Hz, 2H), 7.21 (d, J = 8.4 Hz, 2H), 7.15 (d, J = 1.9 Hz, 2H), 4.73 (s, 4H), 4.69 (s, 2H) , 3.92 (dd, J = 6.0, 2.7 Hz, 4H), 3.72 – 3.65 (m, 4H), 3.31 – 3.29 (m, 2H), 3.08 – 2.87 (m, 4H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 779.18, found: 779.40

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.52-7.35 (m, 10H), 7.16 (d, J = 8.0 Hz, 2H), 7.11 (s, 2H), 7.02-6.82 (m, 2H), 4.74 (s, 4H), 4.11 (m, 4H), 3.49 (m, 4H), 3.02 (t, J = 6 Hz, 4H), 2.90 (m, 2H), 1.81 (m, 2H), 1.69 (m , 2H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.21

¹ H NMR (500 MHz, DMSO-d6) δ 8.20 (s, 1H), 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.8, 1.8 Hz, 2H), 7.39 (dd, J = 7.5, 1.8 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.15 (dd, J = 8.3, 2.0 Hz, 2H), 7.09 (d, J = 2.0 Hz, 2H), 4.92 – 4.87 (m, 2H), 4.69 (s, 4H), 4.03 – 3.95 (m, 4H), 3.55 – 3.51 (m, 6H), 3.29 – 3.27 (m, 2H), 2.85 – 2.80 (m, 2H), 2.71 – 2.64 (m, 2H), 1.16 (t, J = 6.6 Hz, 12H).

LCMS (ESI): for C ₄₄ H ₄₈ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 863.27, found: 863.77

¹ H NMR (500 MHz, DMSO-d6) δ 7.7 (s, 2H), 7.52-7.21 (m, 10H), 4.73 (s, 4H), 4.40-4.20 (m, 4H), 3.94-3.90(m, 2H), 3.50-3.20 (m, 4H), 2.82 (d, J = 4.8 Hz, 4H), 2.87 (m, 2H).

LCMS (ESI): for C ₄₀ H ₃₈ Cl ₂ N ₆ O ₁₀ ; [M+H] ⁺ : calcd: 833.20, found: 833.20

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.5 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.7, 1.8 Hz, 4H), 7.42-7.36 (m, 4H), 7.15 ( dd, J = 8.3, 2.0 Hz, 2H), 4.70 (s, 4H), 4.14 (hept, J = 7.2, 6.7 Hz, 4H), 3.61 (dd, J = 9.6, 4.1 Hz, 2H), 3.54 (dd , J = 9.5, 6.2 Hz, 2H), 3.33 (t, J = 5.1 Hz, 2H), 3.33 (p, J = 6.0 Hz, 4H), 1.10 (s, 18H).

LCMS (ESI): for C ₄₆ H ₅₂ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 891.31, found: 891.31

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 13.07 (s, 2H), 7.52-7.38 (m, 10H), 7.10 (s, 2H), 4.87 (s, 4H), 4.4-4.3 (m, 2H ), 4.22-4.15 (m, 4H), 4.01-3.95 (m, 4H), 3.87-3.79 (m, 4H), 3.11-3.04 (m, 4H).

LCMS (ESI): for C ₄₂ H ₃₆ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 827.18, found: 827.18

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51 (t, J = 7.5Hz, 2H), 7.46 (dd, J = 7.7, 1.9 Hz, 2H), 7.42-7.35 (m, 4H), 7.17 ( dd, J = 8.3, 1.9 Hz, 2H), 7.11 (d, J = 2.0 Hz, 2H), 4.71 (s, 4H), 4.18 (q, J = 11.6, 9.3 Hz, 4H), 3.66 (m, 4H ), 3.26 (s, 6H), 3.07 (m, 4H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.21

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (dd, J = 7.4, 1.9 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.1, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.01 ( t, J = 7.0 Hz, 4H), 3.81 (s, 4H), 3.04 – 3.00 (m, 4H), 1.97 (t, J = 8.1 Hz, 4H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 747.19, found: 747.46

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.51 (t, J = 7.6 Hz, 2H), 7.45 (dd, J = 7.7, 1.8 Hz, 2H), 7.40 (dd, J = 7.4, 1.8 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.3, 2.0 Hz, 2H), 7.13 (d, J = 1.9 Hz, 2H), 4.74 (s, 4H), 4.01 ( t, J = 7.1 Hz, 4H), 3.12 (t, J = 7.0 Hz, 4H), 3.03 (t, J = 7.8 Hz, 4H), 2.61 (d, J = 7.0 Hz, 4H), 1.98 – 1.90 (m, 4H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 775.22, found: 775.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (dd, J = 7.4, 1.9 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.1, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.13 ( t, J = 7.5 Hz, 4H), 4.01 (t, J = 7.0 Hz, 4H), 3.31 – 3.29 (m, 2H), 3.04 – 3.00 (m, 4H), 1.97 (t, J = 8.1 Hz, 4H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (dd, J = 7.4, 1.9 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.1, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 5.47 (s, 2H), 4.73 ( s, 4H), 4.39 – 4.34 (m, 2H), 4.28 – 4.19 (m, 4H), 4.01 (t, J = 7.0 Hz, 4H), 3.71 – 3.67 (m, 2H), 3.24 – 3.20 (m, 2H), 2.68 – 2.56 (m, 2H), 2.46 – 2.34 (m, 2H), 2.20 – 2.11 (m, 4H)

LCMS (ESI): for C ₄₄ H ₄₄ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 859.24, found: 859.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (dd, J = 7.4, 1.9 Hz, 2H), 7.35 (d, J = 8.4 Hz, 2H), 7.18 (dd, J = 8.1, 2.0 Hz, 2H), 7.13 (d, J = 2.0 Hz, 2H), 4.73 (s, 4H), 4.13 ( t, J = 7.5 Hz, 4H), 4.07 (q, J = 7.1 Hz, 4H), 4.01 (t, J = 7.0 Hz, 4H), 3.31 – 3.29 (m, 2H), 3.04 – 3.00 (m, 4H), 1.97 (t, J = 8.1 Hz, 4H), 1.19-1.14 (t, J = 7.1 Hz, 6H).

LCMS (ESI): for C ₄₄ H ₄₈ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : Calculated: 863.27, Found: 863.50 Example 6 Preparation of (2S,2'S)-2,2'-(((2R,2'R)-((2,2'- dichloro- [1,1'- biphenyl ]-3,3' -diyl ) bis (3- oxo -2,3 -dihydro -4H- benzo [b][1,4] oxazine -7,4 -diyl )) bis ( propane -1,2 -diyl )) bis ( azanediyl ) ) bis (3- hydroxypropanoic acid )

Synthesis of intermediate 2

_{A solution of compound SM4 (0.515 g, 2.258 mmol, 1 eq), 1-bromopropan-2-one (0.340 g, 2.48 mmol, 1.10 eq) and Cs2CO3 (} 0.960 g, 2.93 mmol, 1.30 eq) in DMF (5 ml) was stirred at room temperature for 1 hour, 15 mL of _H2O was slowly added, the solid was collected and dried to give the title compound 2 (0.400 g, 62%).

Synthesis of intermediate 4

To a solution of compound SM2 (0.300 g, 1.056 mmol, 1 eq), compound 3 (0.315 g, 2.024 mmol, 1.90 eq), DIEA (0.274 g, 2.124 mmol, 2.01 eq), 4A and 2 drops of AcOH in DCM (6 ml) was added NaBH(OAc) ₃ (1.00 g, 4.73 mmol, 4 eq) at room temperature. After 3 to 4 hours, 10 mL of H ₂ O was added, extracted with DCM, and the organic layer was collected and purified with silica gel to give the title compound 4 (0.07 g, 17%).

Synthesis of intermediate 6

A solution of compound SM2 (0.043 g, 0.09 mmol, 1 eq), compound 4 (0.07 g, 0.18 mmol, 2.0 eq), K ₂ CO ₃ (0.05 g, 0.36 mmol, 4.0 eq), Pd(dppf)Cl ₂ DCM (0.008 g, 0.009 mmol, 0.1 eq) in dioxane (3 ml) and H ₂ O (0.5 ml) was stirred at 80 °C under N ₂ for 2 h, 5 mL of H ₂ O was added, extracted with EA, the organic layer was collected and purified using preparative TLC to give the title compound 6 (0.05 g, 66%).

Synthesis of GLC01-563

To a stirred solution of compound 6 (0.025 g, 0.030 mmol, 1 eq) in MeOH (2 mL) was added a solution of NaOH (0.020 g, 0.50 mmol, 16.6 eq) in _H2O (0.5 mL) at room temperature. After 1 hour, the mixture was purified by reverse phase HPLC (0.1% trifluoroacetic acid in water/acetonitrile) to provide GLC01-563 (12 mg, 50%).

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.49 (d, J = 7.4 Hz, 2H), 7.46 (d, J = 7.4 Hz, 2H), 7.40 (t, J = 8.0 Hz, 4H), 7.14 (d, J = 8.5 Hz, 2H), 7.11 (d, 2H), 4.71 (s, 4H), 4.12 – 4.07 (m, 2H), 4.00 – 3.96 (m, 2H), 3.63 – 3.57 (m, 6H ), 3.24 – 3.22 (m, 2H), 1.08 (d, J = 6.4 Hz, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 807.21, found: 807.43

GLC01-550 was prepared using the same process.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.49 (d, J = 7.4 Hz, 2H), 7.46 (d, J = 7.4 Hz, 2H), 7.40 (t, J = 8.0 Hz, 4H), 7.14 (d, J = 8.5 Hz, 2H), 7.11 (d, 2H), 4.71 (s, 4H), 4.12 – 4.07 (m, 2H), 4.00 – 3.96 (m, 2H), 3.63 – 3.57 (m, 6H ), 3.24 – 3.22 (m, 2H), 1.08 (d, J = 6.4 Hz, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : Calcd: 807.21, Found: 807.43 Example 7 7A. 2'-(((2,2'- dichloro- [1,1' -biphenyl ]-3,3' -diyl ) bis (6- fluoro -3- oxo -2,3- dihydro -4H- benzo [b][1,4] oxazine -7,4 -diyl )) bis ( ethane -2,1 -diyl )) bis ( azanediyl ))(2S,2'S)-bis ( 3- hydroxypropionate ) and ( Preparation of 2S,2'S)-2,2'-((((2,2'- dichloro- [1,1'- biphenyl ]-3,3' -diyl ) bis (6- fluoro -3- oxo -2,3- dihydro -4H- benzo [b][1,4] oxazine -7,4 -diyl )) bis ( ethane -2,1 -diyl )) bis ( azanediyl )) bis (3- hydroxypropionic acid )

Synthesis of intermediate 2

Compound 1 (0.100 g, 0.598 mmol, 1.00 eq), NBS (0.115 g, 0.646 mmol, 1.08 eq), a drop of AcOH in CH ₃ CN (1.5 ml) were stirred at 50° C. for 16 hours, H ₂ O (5 ml) was added slowly, extracted with EA, and the organics were concentrated to give the title compound 2 (0.100 g, 68%).

Synthesis of intermediate 3

A solution of compound 2 (1.50 g, 6.09 mmol, 1.00 eq), 2-bromo-1,1-dimethoxyethane (2.00 g, 11.83 mmol, 1.94 eq) and Cs ₂ CO ₃ (5.00 g, 15.33 mmol, 2.51 eq) in DMF (15 ml) was stirred at 60° C. for 16 hours, 45 mL of H ₂ O was slowly added, and the dried solid was collected in vacuo to obtain compound 3 (0.65 g, 32%).

Synthesis of intermediate 5

A solution of compound SM2 (0.280 g, 0.589 mmol, 1.00 eq), compound 3 (0.400 g, 1.197 mmol, 2.03 eq), K ₂ CO ₃ (0.350 g, 2.536 mmol, 4.3 eq), Pd(dppf)Cl ₂ in DCM (0.030 g, 0.036 mmol, 0.06 eq) in dioxane (3 ml) and H ₂ O (0.5 ml) was stirred at 80 °C under N ₂ for 2 h, 5 mL of H ₂ O was added, extracted with EA, the organic layer was concentrated and purified using silica gel (PE:EA = 3:1-1:1) to give compound 5 (0.330 g, 77%).

Synthesis of intermediate 6

A solution of compound 5 (0.340 g, 0.466 mmol, 1 eq) in dioxane/1 N HCl (4 ml/2 ml) was stirred at 80 ^° C. for 0.5 h, 10 mL of H ₂ O was added, extracted with EA, and the organic layer was concentrated to obtain compound 6 (0.330 g, crude product).

Synthesis of GLC01-589

To a solution of compound 6 (0.330 g, 0.520 mmol, 1.0 eq), compound 7 (0.440 g, 2.59 mmol, 5.0 eq), DIEA (0.351 g, 2.72 mmol, 5.2 eq), 4A and AcOH (0.155 g, 2.59 mmol, 5.0 eq) in DCM (7 ml) was added NaBH(OAc) ₃ (0.500 g, 2.37 mmol, 4.5 eq) at room temperature. After 3 to 4 hours, 10 mL of H ₂ O was added, extracted with DCM, and the organic layer was collected and purified with silica gel (DCM:MeOH = 50:1-20:1) to give GLC01-589 (0.200 g, 44%).

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.53 (t, J = 7.6 Hz, 2H), 7.49 -7.43 (m, 4H), 7.37 (d, J = 11.2 Hz, 2H), 7.04 (d, J = 6.8 Hz, 2H), 4.81 (t, J = 5.6 Hz, 2H), 4.68 (s, 4H), 4.07 (q, J = 7.1 Hz, 4H), 4.00 - 3.90 (m, 4H), 3.60 – 3.49 (m, 4H), 3.37-3.33 (m, 2H), 2.87-2.77 (m, 2H), 2.68 – 2.62 (m, 2H), 1.19-1.14 (t, J = 7.1 Hz, 6H).

LCMS (ESI): for C ₄₂ H ₄₂ Cl ₂ F ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 871.22, found: 871.50

Synthesis of GLC01-554

To a solution of GLC01-589 (0.015 g, 0.017 mmol, 1 eq) in MeOH (0.2 ml) was added NaOH (0.020 g, 0.5 mmol, 29 eq) in _H2O (0.1 ml) at room temperature. After 1 hour, the mixture was purified by reverse phase HPLC (0.1% trifluoroacetic acid in water/acetonitrile) to provide GLC01-554 (7.7 mg, 55%).

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.53 (t, J = 7.6 Hz, 2H), 7.48 – 7.39 (m, 6H), 7.06 (d, J = 6.9 Hz, 2H), 4.70 (s, 4H), 4.13 (t, J = 7.5 Hz, 4H), 3.72 – 3.65 (m, 4H), 3.31 – 3.29 (m, 2H), 3.07 – 3.03 (m, 2H), 3.02 – 2.97 (m, 2H) .

LCMS (ESI): for C ₃₈ H ₃₄ Cl ₂ F ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 815.16, found: 815.33 7B. The following compounds can be prepared using different starting materials.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.54 (m, 2H), 7.49 (d, J = 4 Hz, 2H), 7.43 (d, J = 4.0Hz, 2H), 7.20 (d, J = 8 Hz, 2H), 7.06 (s, 2H), 4.72 (s, 4H), 4.26 (m, 4H), 4.14 (s, 2H), 3.90 (m, 4H), 3.34 (m, 4H).

LCMS (ESI): for C ₃₈ H ₃₄ F ₂ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : Calcd: 815.16, Found: 815.16

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.54 (m, 2H), 7.49 (d, J = 4 Hz, 2H), 7.43 (d, J = 4.0Hz, 2H), 7.30 (d, J = 8 Hz, 2H), 7.06 (s, 2H), 4.72 (s, 4H), 4.26 (m, 4H), 4.14 (s, 2H), 3.90 (m, 4H), 3.34 (m, 4H).

LCMS (ESI): for C ₃₈ H ₃₄ F ₂ Cl ₂ N ₄ O ₁₀ ; [M+H] ⁺ : Calcd: 815.16, Found: 815.16

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.31 (dd, J = 8.1, 5.3 Hz, 4H), 7.21 (d, J = 7.7 Hz, 2H), 7.14 (d, J = 7.5 Hz, 2H) , 7.06 (dd, J = 8.3, 1.7 Hz, 2H), 7.01 (d, J = 2.0 Hz, 2H), 4.82 (s, 2H), 4.68 (s, 4H), 4.07 (q, J = 7.1 Hz, 4H), 4.03-3.90 (m, 4H), 3.56 (hept, J = 5.1 Hz, 4H), 2.85 (dd, J = 12.3 6.8 Hz, 2H), 2.69 (dq, J = 13.3, 7.1, 6.3 Hz, 2H), 1.94 (s, 6H), 1.16 (t, J = 7.0 Hz, 6H),

LCMS (ESI): for C ₄₄ H ₅₀ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 795.35, found: 795.35

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 9.16 (s, 3H), 7.42-7.28 (m, 4H), 7.22 (d, J = 7.7 Hz, 2H), 7.15 (d, J = 7.5 Hz, 2H), 7.10 (dd, J = 8.2, 1.9 Hz, 2H), 7.05 (d, J = 2.0 Hz, 2H), 5.71 (s, 1H), 4.72 (s, 4H), 4.29 (hept, J = 7.5 , 7.0 Hz, 4H), 4.18 (d, J = 3.4 Hz, 2H), 3.91 (d, J = 3.4 Hz, 4H), 3.25 (t, J = 7.7 Hz, 4H), 1.95 (s, 6H),

LCMS (ESI): for C ₄₀ H ₄₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 739.29, found: 739.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.33 (dd, J = 8.3, 5.4 Hz, 4H), 7.22 (d, J = 7.6 Hz, 2H), 7.15 (d, J = 7.5 Hz, 2H) , 7.08 (dd, J = 8.2, 1.8 Hz, 2H), 7.05 (d, J = 2.0 Hz, 2H), 5.47 (s, 2H), 4.72 (s, 4H), 4.39 – 4.34 (m, 2H), 4.28 – 4.19 (m, 4H), 3.71 – 3.67 (m, 2H), 3.24 – 3.20 (m, 2H), 3.08 – 3.02 (m, 2H), 2.68 – 2.56 (m, 2H), 2.46 – 2.34 (m, 2H), 2.20 – 2.11 (m, 4H), 1.95 (s, 6H)..

LCMS (ESI): for C ₄₄ H ₄₆ N ₄ O ₁₀ ; [M+H] ⁺ : Calcd: 791.32, Found: 791.58 Example 8 8A. 2,2'-(((2,2' -difluoro- [1,1' -biphenyl ]-3,3' -diyl ) bis (3- oxo -2,3- dihydro -4H- benzo [b][1,4] oxazine -7,4 -diyl )) bis ( ethane -2,1 -diyl )) bis ( azanediyl ))(2S,2'S)-bis ( 3- hydroxypropionate ) and Preparation of (2S,2'S)-2,2'-((((2,2' -difluoro- [1,1'- biphenyl ]-3,3' -diyl ) bis (3- oxo -2,3- dihydro -4H- benzo [b][1,4] oxazine -7,4 - diyl )) bis ( ethane -2,1 -diyl )) bis ( azanediyl )) bis (3- hydroxypropionic acid )

Synthesis of intermediate GLC01-612-04

To a stirred solution of GLC01-612-03 (100 mg, 0.206 mmol, 1.0 eq) and SM (164 mg, 0.453 mmol, 2.2 eq) in 4 ml of 1,4-dioxane and 0.8 ml of water was added K ₂ CO ₃ (114 mg, 0.826 mmol, 4.0 eq) and Pd(dppf)Cl ₂ ·DCM (33 mg, 0.04 mmol, 0.2 eq) at room temperature under nitrogen atmosphere. The resulting mixture was then heated at 85° C. for 2 hours. Water (15 ml) was added to dilute the reaction mixture and extracted with EA (3 x 15 ml). The combined organic phases were washed with 15 ml of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified using preparative TLC (PE/EA = 2/1) to obtain the desired product (70 mg, yield: 52%) as a light yellow solid.

Synthesis of GLC01-612

To a stirred mixture of GLC01-612-04 (50 mg) in ACN (3 ml) and water (3 ml) was added TFA (0.3 ml) at ambient temperature. After stirring at 80 ^° C for 1 hour, the resulting solution was adjusted to pH = 8 to 9 using aqueous K ₂ CO ₃ solution, extracted with EA, washed with brine, the organic phase was concentrated under reduced pressure, and the residue (43 mg, 100% yield) was used directly in the next step. The above process was followed by a reductive amination step.

¹ H NMR (500 MHz, DMSO) δ 7.53 – 7.31 (m, 8H), 7.12 (dd, J = 31.2, 8.5 Hz, 4H), 4.81 (t, J = 5.5 Hz, 2H), 4.68 (s, 4H ), 4.10 – 4.03 (m, 4H), 3.98 (ddd, J = 21.0, 14.1, 7.1 Hz, 4H), 3.54 (ddt, J = 16.0, 10.5, 5.2 Hz, 4H), 2.83 (dt, J = 14.1 , 7.1 Hz, 2H), 2.69 – 2.63 (m, 2H), 1.22 – 1.12 (m, 6H).

LCMS (ESI): for C ₄₂ H ₄₄ F ₂ N ₄ O ₁₀ ; [M+H] ⁺ : calcd: 803.3, found: 803.3

Synthesis of GLC01-613

The above process is followed by a hydrolysis reaction.

¹ H NMR (500 MHz, DMSO) δ 7.52 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.6, 1.8 Hz, 2H), 7.43 – 7.34 (m, 4H), 7.19 (dd, J = 8.4, 1.9 Hz, 2H), 7.13 (d, J = 1.9 Hz, 2H), 4.73 (s, 4H), 4.31-4.20 (m, 4H), 4.10-4.01 (m, 2H), 3.90-3.81 ( m, 4H), 3.25-3.19 (m, 4H).

LCMS (ESI): for C ₃₈ H ₃₆ F ₂ N ₄ O ₁₀ ; [M+H] ⁺ : Calculated: 747.3, Found: 747.3 Example 9 ( Preparation of Different Cores, Same Side Chains ) 9A. Preparation of 6,6'-(2,2'- dichloro- [1,1'- biphenyl ]-3,3' -diyl ) bis (1-(((S)-5- oxopyrrolidin- 2- yl ) methyl )-3,4- dihydroquinolin - 2(1H) -one )

Substrate A was prepared using the same procedure as for SM5.

A mixture of compound A (100 mg, 1 eq), Cs ₂ CO ₃ (253 mg, 4.0 eq), SM1 (157 mg, 3.0 eq) in DMF was stirred at 40° C. for 2 hours. The reaction mixture was diluted with water and extracted with EA. The combined organic layer was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified using a column (DCM/MeOH = 20/1) to give the title compound 281 (98 mg, yield: 71%). ¹ H NMR (500 MHz, Chloroform- d ) δ 7.46-7.37 (m, 3H), 7.36 (d, J = 2.1 Hz, 1H), 7.33 (dd, J = 7.4, 2.0 Hz, 1H), 7.08 (d, J = 8.5 Hz, 1H), 5.89 (s, 1H) , 4.20-3.98 (m, 3H), 2.99 (t, J = 7.5 Hz, 2H), 2.83-2.66 (m, 2H), 2.40-2.30 (m, 2H), 2.10-1.85 (m, 2H).

LCMS (ESI): for C ₄₀ H ₃₆ Cl ₂ N ₄ O ₄ ; [M+H] ⁺ : calcd: 707.22, found: 707.43 9B. The following compounds can be prepared using different bromide substrates.

¹ H NMR (500 MHz, DMSO-d6) δ 7.93 (s, 2H), 7.80 (d, J = 8.8 Hz, 2H), 7.75 (dd, J = 8.7, 2.1 Hz, 2H), 7.71 (s, 2H ), 7.60 – 7.54 (m, 4H), 7.51 – 7.47 (m, 2H), 3.91 – 3.82 (m, 2H), 3.61 – 3.53 (m, 4H), 3.41 – 3.33 (m, 4H), 2.85 – 2.76 (m, 4H), 2.39 – 2.26 (m, 4H), 2.25 – 2.06 (m, 6H), 1.92 – 1.84 (m, 2H)

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₂ ; [M+H] ⁺ : calcd: 679.25, found: 679.45

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.78 (s, 2H), 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.7, 1.7 Hz, 2H), 7.40 (d, J = 7.4 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 7.14 (d, J = 8.3 Hz, 2H), 7.10 (d, J = 2.0 Hz, 2H), 4.71 (s, 4H) , 4.06 – 3.89 (m, 6H), 2.34 – 2.24 (m, 2H), 2.20 – 2.07 (m, 4H), 1.84 – 1.72 (m, 2H)..

LCMS (ESI): for C ₃₈ H ₃₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 711.17, found: 711.47

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.78 (s, 2H), 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (d, J = 7.5 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 7.14 (dd, J = 8.6, 1.8 Hz, 2H), 7.10 (d, J = 2.1 Hz, 2H), 4.71 (s, 4H), 4.07 – 3.88 (m, 6H), 2.35 – 2.24 (m, 2H), 2.21 – 2.04 (m, 4H), 1.84 – 1.73 (m, 2H)..

LCMS (ESI): for C ₃₈ H ₃₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 711.17, found: 711.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.78 (s, 2H), 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (d, J = 7.5 Hz, 2H), 7.34 (s, 2H), 7.10 (s, 2H), 4.71 (s, 4H), 4.07 – 3.88 (m, 6H), 2.35 – 2.24 (m, 2H), 2.21 – 2.04 (m, 4H), 1.84 – 1.73 (m, 2H).

LCMS (ESI): for C ₃₈ H ₃₄ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 741.19, found: 741.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.29 (s, 1H), 7.93 (t, J = 1.7 Hz, 1H), 7.80 (d, J = 8.9 Hz, 1H), 7.75 (dd, J = 8.7, 2.1 Hz, 1H), 7.70 (s, 1H), 7.57 (d, J = 3.9 Hz, 2H), 7.49 (dd, J = 5.6, 3.8 Hz, 1H), 4.44-4.37 (m, 1H), 4.27-4.20 (m, 1H), 4.11-4.03 (m, 1H), 2.42-2.30 (m, 1H), 2.24-2.16 (m, 1H), 2.03-1.95 (m, 1H), 1.891-1.84 (m, 1H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 705.18, found: 705.52

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.29 (s, 2H), 7.93 (s, 2H), 7.80 (d, J = 8.8 Hz, 2H), 7.75 (dd, J = 8.7, 2.1 Hz, 2H), 7.71 (s, 2H), 7.60 – 7.54 (m, 4H), 7.51 – 7.47 (m, 2H), 4.41 (dd, J = 14.0, 8.6 Hz, 2H), 4.23 (dd, J = 13.8, 5.3 Hz, 2H), 4.09 – 4.02 (m, 2H), 2.60 – 2.54 (m, 2H), 2.41 – 2.32 (m, 3H), 2.25 – 2.06 (m, 5H), 1.92 – 1.84 (m, 2H).

LCMS (ESI): for C ₃₈ H ₃₄ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 709.20, found: 709.44

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.13 (s, 2H), 7.86 (s, 2H), 7.58 – 7.53 (m, 6H), 7.46 (dd, J = 5.9, 3.6 Hz, 2H), 4.21 – 4.15 (m, 2H), 4.05 (dd, J = 13.2, 6.6 Hz, 2H), 4.01 – 3.96 (m, 2H), 2.20 (q, J = 10.8 Hz, 4H), 2.08 (dd, J = 11.3, 3.9 Hz, 4H), 1.53 (s, 6H), 1.48 (s, 6H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 767.24, found: 767.50

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.79-7.71 (m, 2H), 7.55-7.49 (m, 2H), 7.48-7.41 (m, 4H), 7.38 (td, J = 7.2, 1.7 Hz , 2H), 7.06 (d, J = 2.6 Hz, 1H), 7.01 (d, J = 1.9 Hz, 1H), 4.73 (s, 4H), 4.04-3.88 (m, 6H), 2.33-1.94 (m, 6H), 1.78 (d, J = 13.0 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₄ N ₄ O ₆ ; [M+H] ⁺ : calcd: 779.10, found: 779.41

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.94 (s, 2H), 7.62 (d, J = 8 Hz, 2H), 7.56-7.49 (m, 6H), 7.45 (s, 2H), 4.22 ( m, 2H), 4.06 (dd, J = 4 Hz, 2H), 3.95 (m, 2H), 2.32 (m, 2H) , 2.20 (m, 2H) , 2.10 (m, 2H) 1.80 (m, 2H) .

LCMS (ESI): for C ₃₆ H ₂₆ F ₄ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 785.12, found: 785.12

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.13 (s, 2H), 7.86 (s, 2H), 7.58 – 7.53 (m, 6H), 7.46 (dd, J = 5.9, 3.6 Hz, 2H), 4.21 – 4.15 (m, 2H), 4.05 (dd, J = 13.2, 6.6 Hz, 2H), 4.01 – 3.96 (m, 2H), 2.20 (q, J = 10.8 Hz, 4H), 2.08 (dd, J = 11.3, 3.9 Hz, 4H), 1.53 (s, 6H), 1.48 (s, 6H).

LCMS (ESI): for C ₄₀ H ₃₈ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 769.23, found: 769.50

¹ HNMR (500 MHz, DMSO- d ₆ ) δ 7.76 (s, 2H), 7.53 (t, J = 7.4 Hz, 3H), 7.48 – 7.42 (m,4H), 7.31 (t, J = 10.8 Hz, 2H ), 7.05 (d, J = 6.9 Hz, 2H), 4.70 (s, 4H), 4.05 – 3.87 (m, 6H), 2.33 – 2.23 (m, 2H), 2.20 – 2.04 (m, 4H), 1.77 ( dd, J = 13.6, 7.7 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₂ F ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 747.15, found: 747.10

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.79 (d, J = 2.9 Hz, 2H), 7.54 (t, J = 7.5 Hz, 2H), 7.46 (d, J = 7.1 Hz, 4H), 7.19 (d, J = 8.6 Hz, 2H), 7.04 (t, J = 7.8 Hz, 2H), 4.81 (d, J = 14.7 Hz, 4H), 4.08 – 4.01 (m, 2H), 3.93(d, J = 12.0 Hz, 4H), 2.34 – 2.23 (m, 2H), 2.21 – 2.04 (m, 4H), 1.77 (dd, J = 13.2, 8.2 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₂ F ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 747.15, found: 747.11

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.88 (s, 2H), 7.51 (dt, J = 14.3, 7.2 Hz, 4H), 7.42 (d, J = 7.0 Hz, 2H), 7.15 (d, J = 14.0 Hz, 2H), 7.04 (s, 2H), 4.74 – 4.67 (m, 4H), 4.10 (dd, J = 14.3, 7.5 Hz, 2H), 3.98 – 3.85 (m, 4H), 2.19 (t , J = 9.6 Hz, 2H), 2.15 – 2.06 (m, 4H), 1.70 (dd, J = 16.5, 7.0 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₂ F ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 747.15, found: 747.15

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.61 (s, 2H), 7.92 (s, 2H), 7.81 (s, 2H), 7.62-7.75 (m, 6H), 4.78 (s, 4H), 4.22 (m, 2H), 4.06 (dd, J = 4 Hz, 2H), 3.95 (m, 2H), 2.32 (m, 2H) , 2.20 (m, 2H) , 2.10 (m, 2H) 1.80 (m, 2H).

LCMS (ESI): for C ₃₆ H ₃₀ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 713.16, found: 713.16

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.93 (s, 2H), 7.80 (d, J = 8.8 Hz, 2H), 7.75 (dd, J = 8.7, 2.1 Hz, 2H), 7.71 (s, 2H), 7.60 – 7.54 (m, 4H), 7.51 – 7.47 (m, 2H), 4.41 (dd, J = 14.0, 8.6 Hz, 2H), 4.23 (dd, J = 13.8, 5.3 Hz, 2H), 4.09 – 4.02 (m, 2H), 3.50 (s, 6H), 2.60 – 2.54 (m, 2H), 2.41 – 2.32 (m, 3H), 2.25 – 2.06 (m, 5H), 1.92 – 1.84 (m, 2H).

LCMS (ESI): for C ₄₀ H ₃₈ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 737.23, found: 737.52

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.81 (s, 2H), 7.52 (t, J = 7.5 Hz, 2H), 7.46 (dd, J = 7.6, 1.7 Hz, 2H), 7.42 (dd, J = 7.4, 1.9 Hz, 2H), 7.31 (d, J = 8.5 Hz, 2H), 7.25 – 7.20 (m, 2H), 7.19 (d, J = 2.1 Hz, 2H), 5.53 (s, 4H), 3.99 – 3.88 (m, 2H), 3.78 (d, J = 5.6 Hz, 4H), 2.32 – 2.24 (m, 2H), 2.23 – 2.09 (m, 4H), 1.90 – 1.80 (m, 2H).

LCMS (ESI): for C ₃₆ H ₃₆ Cl ₂ N ₄ O ₈ S ₂ ; [M+H] ⁺ : calcd: 783.10, found: 783.42

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.50 (t, J = 7.5 Hz, 1H), 7.48-7.43 (m, 1H), 7.41-7.32 (m, 3H), 7.12 (d, J = 2.1 Hz, 1H), 7.07 (dt, J = 8.3, 1.6 Hz, 1H), 5.38 (s, 1H), 3.91-3.74 (m, 2H), 3.68-3.50 (m, 3H), 2.48 -2.32 (m, 2H), 2.15 -1.93 (m, 4H).

LCMS (ESI): for C ₄₀ H ₃₈ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 737.24, found: 737.57

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.78-7.73 (m, 2H), 7.58 (dd, J = 7.7, 1.9 Hz, 1H), 7.54 (t, J = 7.6 Hz, 1H), 7.43 ( dd, J = 7.4, 1.9 Hz, 1H), 7.38 (d, J = 8.1 Hz, 1H), 4.91 (d, J = 1.4 Hz, 2H), 4.06-4.01 (m, 1H), 3.96-3.90 (m , 2H), 2.37-2.24 (m, 2H), 2.16-2.05 (m, 2H).

LCMS (ESI): for C ₃₆ H ₃₀ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 713.17, found: 713.48

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.71 (s, 2H), 7.92 (s, 2H), 7.62-7.75 (m, 6H), 4.78 (s, 4H), 4.22 (m, 2H), 4.06 (dd, J = 4 Hz, 2H), 3.95 (m, 2H), 2.32 (m, 2H) , 2.20 (m, 2H) , 2.10 (m, 2H) 1.80 (m, 2H).

LCMS (ESI): for C ₃₄ H ₂₈ Cl ₂ N ₈ O ₆ ; [M+H] ⁺ : Calcd: 715.15, Found: 715.15 Example 10 ( Preparation of Same Core, Different Side Chains ) 10A. Preparation of 2,2'-((2,2'- dichloro- [1,1'- biphenyl ]-3,3' -diyl ) bis (3- oxo -2,3 -dihydro -4H- benzo [b][1,4] oxazine -7,4 -diyl )) diacetonitrile

A mixture of compound SM5 (100 mg, 1 eq), Cs ₂ CO ₃ (251 mg, 4.0 eq), bromoacetonitrile (69 mg, 3.0 eq) in DMF was stirred at 40° C. for 2 hours. The reaction mixture was diluted with water and extracted with EA. The combined organic layer was washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The residue was purified using a column (DCM/MeOH = 20/1) to give the title compound 397 (92 mg, yield: 67.6%).

1H NMR (500 MHz, DMSO-d6) δ 7.54-7.46 (m, 2H), 7.44-7.38 (m, 2H), 7.25 (dd, J = 8.3, 2.0 Hz, 1H), 7.18 (d, J = 2.0 Hz, 1H), 5.15 (s, 2H), 4.84 (s, 2H).

LCMS (ESI): for C ₃₂ H ₂₀ Cl ₂ N ₄ O ₄ ; [M+H] ⁺ : calcd: 595.09, found: 595.40 10B. Following the above process, compounds can be prepared from different bromide side chains.

¹ H NMR (500 MHz, DMSO-d6) δ 7.55 (br s, 1H), 7.52-7.45 (m, 2H), 7.41-7.36 (m, 2H), 7.16 (dd, J = 8.4, 2.0 Hz, 1H ), 7.12 (d, J = 2.0 Hz, 1H), 4.73 (s, 2H), 4.11-3.98 (m, 2H), 3.34-3.28 (m, 1H), 3.06-2.99 (m, 1H), 2.83- 2.74 (m, 1H), 2.30-2.22 (m, 1H), 2.02-1.95 (m, 1H).

LCMS (ESI): for C ₃₈ H ₃₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 711.18, found: 711.47

1H NMR (500 MHz, DMSO-d6) δ 8.53 (d, J = 1.2 Hz, 1H), 7.95 (d, J = 1.5 Hz, 1H), 7.52-7.47 (m, 1H), 7.44 (dd, J = 7.7, 1.8 Hz, 1H), 7.39 (dd, J = 7.4, 1.7 Hz, 1H), 7.12-7.08 (m, 3H), 4.67 (s, 2H), 4.49 (t, J = 6.0 Hz, 2H), 4.33 (t, J = 6.1 Hz, 2H).

LCMS (ESI): for C ₃₆ H ₂₈ Cl ₂ N ₈ O ₄ ; [M+H] ⁺ : calcd: 707.17, found: 707.43

1H NMR (500 MHz, DMSO-d6) δ 7.74 (d, J = 1.9 Hz, 1H), 7.52-7.47 (m, 1H), 7.46-7.41 (m, 2H), 7.39 (dd, J = 7.4, 1.8 Hz, 1H), 7.09 (d, J = 1.9 Hz, 1H), 7.06 (dd, J = 8.4, 2.0 Hz, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.18 (t, J = 2.0 Hz, 1H), 4.68 (d, J = 1.4 Hz, 2H), 4.40 (t, J = 6.2 Hz, 2H), 4.28 (t, J = 6.2 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₀ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 705.18, found: 705.42

1H NMR (500 MHz, DMSO-d6) δ 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.7, 1.9 Hz, 2H), 7.42 – 7.36 (m, 4H), 7.15 (dd, J = 8.2, 2.0 Hz, 2H), 7.09 (d, J = 2.0 Hz, 2H), 4.93 (t, J = 5.7 Hz, 2H), 4.69 (s, 4H), 4.00 (t, J = 6.1 Hz, 4H), 3.65 – 3.59 (m, 4H)..

LCMS (ESI): for C ₃₂ H ₂₆ Cl ₂ N ₂ O ₆ ; [M+H] ⁺ : Calcd: 605.12, Found: 605.50

1H NMR (500 MHz, DMSO-d6) δ 7.52-7.45 (m, 2H), 7.41-7.37 (m, 2H), 7.18 (dd, J = 8.4, 2.0 Hz, 1H), 7.10 (d, J = 2.0 Hz, 1H), 4.68 (s, 2H), 4.06 (t, J = 6.5 Hz, 2H), 3.43 (dd, J = 9.0, 6.8 Hz, 2H), 3.31 (t, J = 6.5 Hz, 2H), 3.17 (dd, J = 8.9, 6.8 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₄ Cl ₂ N ₆ O ₆ ; [M+H] ⁺ : calcd: 741.20, found: 741.50

1H NMR (500 MHz, DMSO-d6) δ 7.48 (t, J = 7.6 Hz, 1H), 7.43 (dd, J = 7.6, 1.9 Hz, 1H), 7.35 (dd, J = 7.4, 1.8 Hz, 1H) , 7.07 (dd, J = 8.3, 2.0 Hz, 1H), 7.03 (d, J = 2.0 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 4.65 (s, 2H), 4.16 (s, 2H).

LCMS (ESI): for C ₃₂ H ₂₂ Cl ₂ N ₂ O ₈ ; [M+H] ⁺ : calcd: 633.08, found: 633.37

1H NMR (500 MHz, DMSO-d6) δ 8.87 (d, J = 2.0 Hz, 2H), 8.74 (d, J = 2.1 Hz, 2H), 8.28 (d, J = 2.3 Hz, 2H), 7.52 (t , J = 7.6 Hz, 2H), 7.46 (dd, J = 7.7, 1.9 Hz, 2H), 7.44 – 7.39 (m, 4H), 7.16 (dd, J = 8.3, 2.0 Hz, 2H), 7.12 (d, J = 2.0 Hz, 2H), 4.67 (s, 4H), 4.23 (t, J = 7.3 Hz, 4H), 3.01 (t, J = 7.3 Hz, 4H).

LCMS (ESI): for C ₄₄ H ₃₀ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 777.17, found: 777.45

1H NMR (500 MHz, DMSO-d6) δ 7.52 (t, J = 7.6 Hz, 2H), 7.46 (d, J = 7.8 Hz, 2H), 7.41 (d, J = 7.5 Hz, 2H), 7.36 (d , J = 8.4 Hz, 2H), 7.19 (d, 2H), 7.14 (d, J = 2.0 Hz, 2H), 4.75 (s, 4H), 3.75 – 3.67 (m, 4H), 3.24 – 3.17 (m, 4H), 3.08 – 3.02 (m, 4H), 2.34 – 2.16 (m, 4H), 1.97 – 2.01(m, 4H).

LCMS (ESI): for C ₄₀ H ₃₆ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 739.20, found: 739.50

1H NMR (500 MHz, DMSO-d6) δ 7.54-7.48 (m, 2H), 7.46 (dd, J = 7.6, 1.9 Hz, 1H), 7.40 (dd, J = 7.4, 1.9 Hz, 1H), 7.19 ( dd, J = 8.4, 2.1 Hz, 1H), 7.13 (d, J = 2.0 Hz, 1H), 4.59 (s, 2H), 4.52 (d, J = 13.1 Hz, 1H), 4.39 (t, J = 12.3 Hz, 1H), 3.92 (d, J = 13.5 Hz, 1H), 3.25-3.13 (m, 1H), 2.70-2.53 (m, 2H), 2.42-2.27 (m, 1H), 2.04 (s, 3H), 1.86-1.71 (m, 2H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 767.24, found: 767.56

1H NMR (500 MHz, DMSO-d6) δ 7.53-7.31 (m, 4H), 7.25 (dd, J = 8.3, 2.0 Hz, 1H), 7.05 (d, J = 2.0 Hz, 1H), 5.36 (s, 2H), 4.77 (s, 2H).

LCMS (ESI): for C ₃₂ H ₂₂ Cl ₂ N ₁₀ O ₄ ; [M+H] ⁺ : calcd: 681.13, found: 681.41

1H NMR (500 MHz, DMSO-d6) δ 7.84 (s, 1H), 7.51 (t, J = 7.5 Hz, 1H), 7.46 (dd, J = 7.7, 1.9 Hz, 1H), 7.42-7.38 (m, 1H), 7.36 (d, J = 8.5 Hz, 1H), 7.15 (dd, J = 8.3, 2.1 Hz, 1H), 7.11 (d, J = 2.0 Hz, 1H), 4.72 (d, J = 3.3 Hz, 2H), 4.41 (t, J = 7.9 Hz, 1H), 4.20-4.08 (m, 3H), 4.02-3.94 (m, 1H).

LCMS (ESI): for C ₃₆ H ₂₈ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 715.14, found: 715.42

1H NMR (500 MHz, DMSO-d6) δ 7.78 (s, 2H), 7.50 (t, J = 7.5 Hz, 2H), 7.45 (dd, J = 7.8, 1.8 Hz, 2H), 7.40 (d, J = 7.5 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 7.14 (dd, J = 8.6, 1.8 Hz, 2H), 7.10 (d, J = 2.1 Hz, 2H), 7.09 (d, J = 5.4, 2.8 Hz, 2H), 6.96 (d, J = 2.7 Hz, 2H), 4.74 (s, 4H),

LCMS (ESI): for C ₃₆ H ₂₆ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 677.14, found: 677.43

1H NMR (500 MHz, DMSO-d6) δ 8.95 (d, J = 2.0 Hz, 1H), 8.91 (d, J = 2.3 Hz, 1H), 8.29 (q, J = 2.4 Hz, 1H), 7.47 (dq , J = 7.7, 4.2 Hz, 1H), 7.42 (dt, J = 7.5, 2.4 Hz, 1H), 7.37 (dt, J = 7.6, 2.3 Hz, 1H), 7.13 (d, J = 4.1 Hz, 1H) , 7.05 (d, J = 4.2 Hz, 2H), 5.27 (d, J = 4.2 Hz, 2H), 4.91 (d, J = 4.2 Hz, 2H).

LCMS (ESI): for C ₄₂ H ₂₆ Cl ₂ N ₆ O ₄ ; [M+H] ⁺ : calcd: 749.15, found: 749.44 Example 11 ( same core, different side chains ) 11A. Preparation of 7,7'-(2,2'- dichloro- [ 1,1'- biphenyl ]-3,3' -diyl ) bis (4-(((S) -pyrrolidin- 2 -yl ) methyl )-2H- benzo [b][1,4] oxazin -3(4H) -one )

1.

To a stirred solution of 411-01 (500 mg, 2.49 mmol, 1.0 eq) in DCM (5 ml) was added TosCl (567 mg, 2.98 mmol, 1.2 eq) and TEA (502 mg, 4.97 mmol, 2.0 eq), then DMAP (30 mg, 0.249 mmol, 0.1 eq) was added at room temperature and stirred for 4 hours. Water and DCM were added to the reaction and separated, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was used directly in the next step (950 mg, yield: 107%).

2.

To a stirred solution of SM4 (117 mg, 0.51 mmol, 1.0 eq) in DMF (2 ml) were added 411-02 (219 mg, 0.62 mmol, 1.2 eq) and CS ₂ CO ₃ (337 mg, 1.03 mmol, 2.0 eq) at ambient temperature. The resulting mixture was then stirred at 70° C. overnight. The reaction was quenched with water and extracted with EA. The combined organic phases were dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated in vacuo. The residue was purified using preparative TLC (PE/EA = 4/1) to give the desired product (105 mg, yield: 50%) as a light yellow oil.

3.

Under _N2 atmosphere, Pd(dppf)Cl2·DCM (19 mg, 0.024 mmol, 0.2 eq), K2CO3 (64 mg, 0.48 mmol, 4.0 eq) were added to a solution of 411-03 (105 mg, 0.26 mmol, 2.2 eq) and (55 mg, 0.12 mmol, 1.0 eq) _SM2 in dioxane (2.5 ml) and _H2O (0.5 ml) under _N2 atmosphere. The final mixture was then heated at 80 _° C for stirring for 2 hours. Cooled to room temperature, water was added and extracted twice with EA, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified using preparative TLC (DCM/MeOH = 15/1) to give the desired product (50 mg, yield: 49%) as a white solid.

4.

To a stirred solution of 411-03 (50 mg, 0.056 mmol, 1.0 eq) in MeOH (1 ml) was added a solution of HCl in dioxane (1 ml, 4 M), and the solution was stirred at room temperature for 2 hours. After the reaction was completed, the solvent was removed by reduced pressure and dried under lyophilization to obtain the desired product (36 mg, yield: 95%).

1H NMR (500 MHz, DMSO-d6) δ 9.61 (s, 2H), 8.72 (s, 2H), 7.52 (t, J = 7.5 Hz, 2H), 7.48 – 7.43 (m, 4H),7.41 (d, J = 7.4 Hz, 2H), 7.19 – 7.12 (m, 4H), 4.78 (t, J = 8.6 Hz, 4H), 4.35 (dd, J = 15.0, 7.8 Hz, 2H), 4.24 (dd, J = 14.9 , 5.0 Hz, 2H), 3.71 (s, 2H), 3.28 (s, 2H), 3.11 (s, 2H), 2.17 (d, J = 6.7 Hz, 2H), 2.02 – 1.88 (m, 4H), 1.71 (dt, J = 18.8, 9.3 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₄ (free form); [M+H] ⁺ : calcd: 683.2, found: 683.2 11B. The following compounds can also be prepared:

¹ H NMR (500 MHz, DMSO-d6) δ 9.26 (s, 4H), 7.51 (t, J = 7.5 Hz, 2H), 7.48 – 7.42 (m, 4H), 7.40 (d, J =7.3 Hz, 2H ), 7.17 (d, J = 8.6 Hz, 2H), 7.13 (s, 2H), 4.73 (d, J = 12.0 Hz, 4H), 4.08 (ddd, J = 41.2, 14.4,7.4 Hz, 4H), 3.32 – 3.23 (m, 4H), 3.15 – 3.05 (m, 2H), 2.98 – 2.89 (m, 2H), 2.66 (dd, J = 15.2, 7.7 Hz, 2H), 2.07 – 1.99 (m, 2H), 1.69 (dq, J = 16.8, 8.4 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₄ (isolated form); [M+H] ⁺ : calcd: 683.2, found: 683.2

1H NMR (500 MHz, DMSO-d6) δ 10.35 (s, 2H), 9.63 (s, 2H), 7.54 – 7.45 (m, 6H), 7.41 (d, J = 7.3 Hz, 2H), 7.16 (d, J = 8.6 Hz, 2H), 7.14 (s, 2H), 4.81 – 4.72 (m, 4H), 4.45 (dd, J = 15.2, 7.8 Hz, 2H), 4.28 (dd, J =15.2, 4.7 Hz, 2H ), 4.07 (s, 2H), 3.84 (dd, J = 24.5, 11.5 Hz, 2H), 3.73 – 3.54 (m, 4H), 2.92 (d, J = 14.7 Hz, 2H).

LCMS (ESI): calculated for C ₃₈ H ₃₂ Cl ₂ F ₄ N ₄ O ₄ (isolated form); [M+H] ⁺ : calculated: 755.2, found: 755.2

1H NMR (500 MHz, DMSO-d6) δ 9.25 (s, 4H), 7.51 (t, J = 7.5 Hz, 2H), 7.48 – 7.42 (m, 4H),7.40 (d, J = 7.3 Hz, 2H) , 7.17 (d, J = 8.5 Hz, 2H), 7.13 (s, 2H), 4.74 (s, 4H), 4.08 (ddd, J = 41.0, 14.5, 7.4 Hz, 4H), 3.31 – 3.22 (m, 4H ), 3.11 (dd, J = 11.9, 5.8 Hz, 2H), 2.94 (td, J = 13.5, 6.5 Hz, 2H), 2.66 (dd, J = 15.2,7.6 Hz, 2H), 2.04 (dt, J = 12.6, 6.4 Hz, 2H), 1.69 (dq, J = 16.7, 8.3 Hz, 2H).

LCMS (ESI): calculated for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₄ (isolated form); [M+H] ⁺ : calculated: 683.2, found: 683.2

1H NMR (500 MHz, DMSO-d6) δ 9.72 (s, 2H), 8.51 (d, J = 9.4 Hz, 2H), 7.52 (t, J = 7.5 Hz, 2H), 7.47 – 7.40 (m, 6H) , 7.16 (d, J = 8.5 Hz, 2H), 7.14 (s, 2H), 4.77 (s, 4H), 4.42 (dd, J = 15.4, 8.2 Hz, 2H), 4.19 (dd, J = 15.0, 4.3 Hz, 2H), 3.75 (s, 2H), 3.51 (s, 2H), 2.21 (d, J = 8.5 Hz, 2H), 2.14 – 2.06 (m, 2H), 1.83 (dd, J = 13.9, 8.4 Hz, 2H), 1.67 – 1.59 (m, 2H), 1.33 (d, J = 6.4 Hz, 6H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₄ (isolated form); [M+H] ⁺ : calcd: 711.2, found: 711.2

1H NMR (500 MHz, DMSO-d6) δ 9.14 (s, 2H), 8.87 (s, 2H), 7.52 (t, J = 7.5 Hz, 2H), 7.47 (d, J = 7.4 Hz, 2H), 7.43 – 7.38 (m, 4H), 7.19 (d, J = 8.3 Hz, 2H), 7.15 (s, 2H), 4.77 (dd, J = 31.2, 15.1 Hz, 4H), 4.39 – 4.30 (m, 2H), 4.03 (s, 2H), 3.91 (d, J = 14.2 Hz, 2H), 3.48 (s, 2H), 1.95 (d, J = 10.9 Hz, 2H), 1.73 (dd, J = 18.0, 10.2 Hz, 2H), 1.62 (dd, J = 18.2, 9.8 Hz, 6H), 1.33 (d, J = 8.7 Hz, 2H), 1.23 (s, 2H ).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₄ (isolated form); [M+H] ⁺ : calcd: 735.2, found: 735.2

1H NMR (500 MHz, DMSO-d6) δ 9.82 (s, 2H), 9.03 (s, 2H), 7.53 – 7.46 (m, 6H), 7.41 (d, J = 7.5 Hz, 2H), 7.17 (d, J = 9.3 Hz, 2H), 7.14 (s, 2H), 4.78 (d, J = 15.5 Hz, 4H), 4.41 – 4.30 (m, 4H), 3.94 (s, 2H), 3.22 (s, 2H), 3.11 – 3.05 (m, 2H), 2.02 – 1.92 (m, 4H), 0.74 – 0.61 (m, 8H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₄ (isolated form); [M+H] ⁺ : calcd: 735.2, found: 735.2

1H NMR (500 MHz, DMSO-d6) δ 8.21 (s, 6H), 7.74 (d, J = 8.4 Hz, 2H), 7.54 – 7.45 (m, 4H), 7.40 (d, J = 7.3 Hz, 2H) , 7.16 (d, J = 7.9 Hz, 2H), 7.11 (s, 2H), 4.72 (dd, J = 39.8, 15.0 Hz, 4H), 4.45 (s, 2H), 4.27 – 4.19 (m, 2H), 3.95 (d, J = 4.6 Hz, 2H), 3.80 (d, J = 9.3 Hz, 2H), 3.68 – 3.56 (m, 4H), 2.11 (d, J = 5.7 Hz, 4H), 1.97 – 1.89 (m, 2H), 1.83 (s, 4H), 1.00 (d, J = 6.8 Hz, 6H), 0.91 (d, J = 6.7 Hz, 6H ).

LCMS (ESI): for C ₄₈ H ₅₄ Cl ₂ N ₆ O ₆ (isolated form); [M+H] ⁺ : calcd: 881.4, found: 881.4

1H NMR (500 MHz, DMSO-d6) 7.73 (d, J = 8.4 Hz, 2H), 7.53 – 7.44 (m, 4H), 7.39 (d, J = 7.3 Hz, 2H), 7.15 (d, J = 7.9 Hz, 2H), 7.09 (s, 2H), 4.72 (m, 4H), 4.44 (s, 2H), 4.27 – 4.19 (m, 2H), 3.95 (d, J = 4.6 Hz, 2H), 3.80 (d , J = 9.3 Hz, 2H), 3.70 (s, 6H), 3.68 – 3.56 (m, 4H), 2.11 (d, J = 5.7 Hz, 4H), 1.97 – 1.89 (m, 2H), 1.83 (s, 4H), 1.00 (d, J = 6.8 Hz, 6H), 0.91 (d, J = 6.7 Hz, 6H).

LCMS (ESI): for C ₅₂ H ₅₈ Cl ₂ N ₆ O ₁₀ ; [M+H] ⁺ : calcd: 997.4, found: 997.4

1H NMR (500 MHz, DMSO-d6) δ 9.74 (s, 2H), 8.84 (s, 2H), 7.53 – 7.44 (m, 6H), 7.41 (d, J = 7.2 Hz, 2H), 7.18 (d, J = 8.5 Hz, 2H), 7.14 (s, 2H), 5.32 (s, 2H), 4.76 (t, J = 10.6 Hz, 4H), 4.35 (ddd, J = 20.6, 17.7, 8.9 Hz, 8H), 3.91 (d, J = 5.5 Hz, 2H), 3.00 – 2.93 (m, 2H), 2.09 (dd, J = 12.9, 5.0 Hz, 2H), 1.87 (td, J = 12.8, 4.7 Hz, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₆ (isolated form); [M+H] ⁺ : calcd: 715.2, found: 715.2

1H NMR (500 MHz, DMSO-d6) δ 7.78 (d, J = 8.4 Hz, 2H), 7.53 – 7.44 (m, 4H), 7.40 (d, J = 7.3 Hz, 2H), 7.17 (d, J = 8.2 Hz, 2H), 7.10 (s, 2H), 4.78 – 4.64 (m, 4H), 4.24 (dd, J = 24.9, 11.7 Hz, 4H), 3.74 (d, J = 10.6 Hz, 2H), 3.52 ( t, J = 9.0 Hz, 2H), 3.38 (dd, J = 17.3, 9.7 Hz, 2H), 2.09 – 2.01 (m, 2H), 1.94 (s, 6H), 1.92 – 1.87 (m, 2H), 1.80 – 1.70 (m, 4H).

LCMS (ESI): for C ₄₂ H ₄₀ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 767.2, found: 767.2

1H NMR (500 MHz, DMSO-d6) δ 9.65 (br s, 1H), 8.76 (br s, 1H), 7.54-7.49 (m, J = 7.6 Hz, 1H), 7.46 (dd, J = 9.0, 3.1 Hz, 2H), 7.41 (d, J = 7.3 Hz, 1H), 7.16 (dd, J = 8.2, 2.1 Hz, 1H), 7.13 (d, J = 2.0 Hz, 1H), 4.77 (s, 2H), 4.38-4.25 (m, 2H), 3.70 (s, 1H), 3.31-3.04 (m, 2H), 2.20-1.95 (m, 2H), 1.94-1.66 (m, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₄ ; [M+H] ⁺ : calcd: 683.22, found: 683.49

1H NMR (500 MHz, DMSO-d6) δ 9.07 (br s, 1H), 8.81 (br s, 1H), 7.52 (m, 1H), 7.48-7.44 (m, 2H), 7.41 (dd, J = 7.5 , 1.8 Hz, 1H), 7.18-7.10 (m, 2H), 4.77 (s, 2H), 4.35-4.01 (m, 2H), 3.30-3.31 (m, 2H), 2.84 (s, 1H), 2.02- 1.91 (m, 2H), 1.73-1.61 (m, 2H), 1.56-1.42 (m, 2H).

LCMS (ESI): for C ₄₀ H ₄₀ Cl ₂ N ₄ O ₄ ; [M+H] ⁺ : calcd: 711.25, found: 711.43

1H NMR (500 MHz, DMSO-d6) δ 9.74 (s, 1H), 9.40 (s, 1H), 7.51 (d, J = 7.9 Hz, 2H), 7.47 (d, J = 1.8 Hz, 1H), 7.41 (d, J = 7.5 Hz, 1H), 7.15 (d, J = 8.4 Hz, 1H), 7.12 (d, J = 1.9 Hz, 1H), 4.77 (d, J = 2.2 Hz, 2H), 4.36-4.25 (m, 1H), 4.16-4.04 (m, 2H), 3.91-3.74 (m, 2H), 3.69-3.53 (m, 2H), 3.32-3.26 (m, 2H).

LCMS (ESI): for C ₃₈ H ₃₆ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 715.21, found: 715.39

1H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 6H), 7.52 (t, J = 6 Hz, 2H), 7.45 (d, J = 4.0 Hz, 2H), 7.40 (d, J = 4 Hz , 2H), 7.35 (d, J = 4 Hz, 2H), 7.17 (d, J = 4.0 Hz, 2H), 7.13 (s, 2H), 4.73 (s, 4H), 4.19 (t, J = 6 Hz , 4H), 3.09 (t, J = 6 Hz, 4H).

LCMS (ESI): for C ₃₂ H ₂₈ Cl ₂ N ₄ O ₄ ; [M+H] ⁺ : Calculated: 603.15, Found: 603.15 Example 12 ( Asymmetric ) Preparation of (S)-7-(2,2'- dichloro -3'-(3- oxo -3,4- dihydro -2H- benzo [b][1,4] oxazin - 7- yl )-[1,1'- biphenyl ]-3- yl )-4-((5 -oxopyrrolidin -2- yl ) methyl )-2H- benzo [b][1,4] oxazin -3(4H) -one

SM5 (20.0 mg, 1 eq), SM1 (10.4 mg, 1 eq) and cesium carbonate (25 mg, 2 eq) were dissolved in DMF (1 ml). The reaction was carried out at 40°C for 2 hours. After cooling, 5 ml of water and 5 ml of EA were added for extraction, and the organic phase was washed with water and concentrated to dryness. The residue was purified using a column (DCM/MeOH = 20/1) to obtain the title compound 429 (11 mg, yield: 46.3%). ¹ H NMR (500 MHz, DMSO- d ₆ ) δ 10.82 (s, 1H), 7.76 (s, 1H), 7.52-7.46 (m, 2H), 7.46-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.34 (d, J = 8.4 Hz, 1H ), 7.14 (dd, J = 8.3, 2.1 Hz, 1H), 7.10 (d, J = 2.0 Hz, 1H), 7.07-7.02 (m, 2H), 6.98 (d, J = 7.9 Hz, 1H), 4.71 (s, 2H), 4.63 (s, 2H), 4.04-3 .92 (m, 3H), 2.34-2.23 (m, 1H), 2.21-2.05 (m, 2H), 1.83-1.74 (m, 1H).LCMS(ESI): for C ₃₃ H ₂₅ Cl ₂ N ₃ O ₅ ; [M+H] ⁺ : calcd: 614.13, found: 614.41

After the above process, the compound designated as title 385 was obtained by the same process.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 10.82 (s, 1H), 7.52-7.45 (m, 3H), 7.43 (dd, J = 7.7, 1.8 Hz, 1H), 7.40-7.36 (m, 3H ), 7.17 (dd, J = 8.3, 2.0 Hz, 1H), 7.10 (d, J = 2.0 Hz, 1H), 7.07-7.02 (m, 2H), 6.98 (d, J = 7.9 Hz, 1H), 4.68 (s, 2H), 4.62 (s, 2H), 4.05 (t, J = 6.5 Hz, 2H), 3.46-3.41 (m, 2H), 3.31 (t, J = 6.5 Hz, 2H), 3.19-3.14 (m, 2H).LCMS(ESI): for C ₃₃ H ₂₆ Cl ₂ N ₄ O ₅ ; [M+H] ⁺ : Calculated value: 629.14, Measured value: 629.44 Example 13 13A. (2-(7-(2,2'- dichloro -3'-(3- oxo -4-(((S) -pyrrolidin -2- yl ) methyl )-3,4- dihydro -2H- benzo [b][1,4] oxazin -7- yl )-[1,1'- biphenyl ]-3- yl )-3- oxo -2 Preparation of 3- dihydro -4H- benzo [b][1,4] oxazin -4- yl ) ethyl )-L- serine

(a) Referring to the following reaction formula, compound 1A-1 (0.35 g, 1.54 mmol, 1 eq), compound 1A-2 (1.10 g, 3.07 mmol, 2.0 eq) and cesium carbonate (0.75 g, 2.30 mmol, 1.5 eq) were dissolved in DMF (10 ml). The reaction was carried out at 60° C. for 5 hours. After cooling, 10 ml of water and 10 ml of EA (ethyl acetate) were added for extraction, and the organic phase was washed with water and purified on silica gel, eluted with PE/EA (v/v, 8:1) to obtain compound 1A, (S)-2-((7-bromo-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester. (0.35 g, yield: 55.5%)

(b) Referring to the following reaction equation, compound 1B (25 mg) was dissolved in dioxane (0.5 ml), and then 1 N HCl solution (0.5 ml) was added dropwise, the temperature was raised to 85°C, and the reaction was carried out for 0.5 hours. The pH value of the reaction solution was adjusted to 7 to 8 by adding a saturated Na ₂ CO ₃ solution, and 10 ml of EA was added for extraction. The organic phase was concentrated to obtain 1B (25 mg, yield: 107.0%).

(c) Referring to the following reaction equation, compound 1C (25 mg, 0.09 mmol, 1 eq), 1D (29 mg, 0.19 mmol, 2 eq), TEA (19 mg, 0.19 mmol) and a drop of AcOH were dissolved in DCM (3 ml), and the resulting mixture was stirred at room temperature for 1 hour. Then, NaBH(OAc) ₃ (37 mg, 0.45 mmol, 5 eq) was added and reacted for 1 hour. 10 ml of H ₂ O and 10 ml of DCM were added for extraction. The organic phase was concentrated to obtain 1C (25 mg, yield: 72%).

(d) Referring to the following reaction equation, compound 1F (0.05 g, 0.11 mmol, 1 eq), 1A (0.043 g, 0.11 mmol, 1 eq), 1E (0.04 g, 0.11 mmol, 1 eq), Pd(dppf)Cl ₂ (0.008 g, 0.01 mmol, 0.1 eq) and potassium carbonate (0.058 g, 0.42 mmol, 4 eq) were dissolved in dioxane/H ₂ O (3 ml, v/v = 5:1). The reaction was carried out at 85° C. for 2 hours under N ₂ atmosphere. After cooling, 10 ml of water and 10 ml of EA were added for extraction, and the organic phase was concentrated and purified using preparative TLC (elution with PE/EA = 1/1) to obtain Compound 1G, (S)-2-((7-(2,2'-dichloro-3'-(4-(2-(((S)-3-hydroxy-1-methoxy-1-oxopropan-2-yl)amino)ethyl)-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-7-yl)-[1,1'-biphenyl]-3-yl)-3-oxo-2,3-dihydro-4H-benzo[b][1,4]oxazin-4-yl)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester, 0.017 g, yield: 20%.

LCMS (ESI): for C ₄₄ H ₄₆ Cl ₂ N ₄ O ₉ ; [M+H] ⁺ : calcd: 845.26, found: 845.26.

(e) Referring to the following reaction equation, 1N HCl (1 mL) was added to a solution of compound 1G (17 mg, 0.02 mmol) in dioxane (1 mL), followed by stirring at room temperature for 3 hours. Then, Na ₂ CO ₃ (aqueous) was added to the reaction mixture to neutralize unreacted HCl. The reaction mixture was extracted with EA and water. The organic phase was concentrated and purified using preparative HPLC to obtain 1F (10 mg, yield: 67%).

(e) Referring to the following reaction equation, NaOH (0.5 ml, 0.26 M in water) was added to a solution of 1F (10 mg, 0.013 mmol) in MeOH/THF (3 mL, 2:1), followed by stirring at room temperature for 1 hour. Then, HCl (1 M) was added to the reaction mixture to neutralize unreacted NaOH. The reaction mixture was purified using preparative HPLC to obtain the title compound 1 (7.4 mg, yield: 75.5%).

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.08 (s, 1H), 8.43 (s, 1H), 7.53-7.36 (m, 8H), 7.19-7.12 (m, 4H), 5.58 (s, 1H ), 4.78 (s, 2H), 4.71 (s, 2H), 4.36 (m, 1H), 4.32-4.13 (m, 3H), 4.04 (m, 1H), 3.87 (s, 2H), 3.72 (m, 1H), 3.21 (m, 2H), 3.12 (m, 2H), 2.20 (m, 1H), 1.94 (m, 2H), 1.70 (m, 1H).

LCMS (ESI): for C ₄₀ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 731.21, found: 731.21

13B. Use the same procedure to prepare the following compounds.

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.09 (s, 1H), 8.64 (s, 2H), 8.44 (s, 1H), 8.15 (t, J = 6.0 Hz, 1H), 7.52 (t, J = 8.0 Hz, 2H), 7.45 (d, J = 4.0 Hz, 2H), 7.41 (d, J = 8.0 Hz, 2H), 7.36 (m, 2H), 7.17 (d, J = 8.0 Hz, 2H) , 7.14 (s, 2H), 4.75 (s, 2H), 4.71 (s, 2H), 4.36 (m, 1H), 4.24 (t, J = 4.0 Hz, 2H), 419 (dd, J = 12.0 Hz, 4.0 Hz, 1H), 3.72 (m, 1H), 3.31 (m, 3H), 3.23 (m, 2H), 3.12 (m, 1H), 3.06 (m, 2H), 2.20 (m, 1H), 1.97 (m, 2H), 1.85 (s, 3H), 1.70 (m, 1H)

LCMS (ESI): for C ₃₉ H ₃₉ Cl ₂ N ₅ O ₅ ; [M+H] ⁺ : calcd: 728.23, found: 728.23

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.09 (s, 1H), 8.44 (s, 1H), 7.53-7.49 (m, 2H), 7.47-7.43 (m, 2H), 7.42-7.33 (m , 4H), 7.21-7.12 (m, 3H), 7.10 (s, 1H), 5.54, (s, 1H), 4.75 (s, 2H), 4.71 (s, 2H), 4.46-4.40 (m, 2H) , 4.34-4.28 (m, 2H), 4.06-3.89 (m, 3H), 3.78-3.66 (m, 2H), 3.44-3.40 (m, 2H), 3.27-3.03 (m, 2H), 2.32-2.20 (m, 2H), 2.18-2.08 (m, 2H), 2.02-1.91 (m, 2H), 1.75-1.86 (m, 2H).

LCMS (ESI): for C ₃₉ H ₃₈ Cl ₂ N ₄ O ₅ ; [M+H] ⁺ : calcd: 713.22, found: 713.22

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 10.84 (s, 1H), 7.51-7.37 (m, 7H), 7.17-6.97 (m, 5H), 4.71 (s, 2H), 4.62 (s, 2H ), 4.15 (t, J = 4.0 Hz, 2H), 3.70-3.62 (m, 3H), 3.05-2.99 (m, 2H).

LCMS (ESI): for C ₃₃ H ₂₇ Cl ₂ N ₃ O ₇ ; [M+H] ⁺ : Calcd: 648.12, Found: 648.12

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.01 (s, 1H), 7.53-7.48 (m, 2H), 7.47-7.44 (m, 2H), 7.41-7.38 (m, 2H), 7.36-7.33 (m, 2H), 7.18 (d, J = 4.0 Hz, 1H), 7.15-7.12 (m, 2H), 7.10 (s, 1H), 4.75 (s, 2H), 4.71 (s, 2H), 4.22 ( t, J = 4.0 Hz, 2H), 4.05-4.00 (m, 1H), 3.96-3.93 (m, 2H), 3.69-3.58 (m, 1H), 3.29 (t, J = 4.0 Hz, 2H), 3.27-3.25 (m, 3H), 2.16-2.06 (m, 2H), 2.30-2.28 (m, 2H), 2.12-2.02 (m, 2H) .

LCMS (ESI): for C ₄₀ H ₃₆ Cl ₂ N ₄ O ₈ ; [M+H] ⁺ : calcd: 771.19, found: 771.19

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 8.61 (s, 2H), 8.14 (t, J = 4.0 Hz, 1H), 7.78 (s, 1H), 7.53-7.48 (m, 2H), 7.47- 7.44 (m, 2H), 7.41-7.38 (m, 2H), 7.36-7.33 (m, 2H), 7.18 (d, J = 4.0 Hz, 1H), 7.15-7.12 (m, 2H), 7.10 (s, 1H), 4.75 (s, 2H), 4.71 (s, 2H), 4.24 (t, J = 4.0 Hz, 2H), 4.04-4.00 (m, 1H), 3.97-3.92 (m, 2H), 3.32-3.30 (t, J = 4.0 Hz, 2H), 3.25-3.21 (m, 2H), 3.08-3.04 (m, 2H), 2.31-2.26 (m, 2H), 2.17-2.07 (m, 2H), 1.85 (s, 3H).

LCMS (ESI): for C ₃₉ H ₃₇ Cl ₂ N ₅ O ₆ ; [M+H] ⁺ : calcd: 742.21, found: 742.21

¹ H NMR (400 MHz, DMSO- d ₆ ) δ 7.78 (s, 1H), 7.53-7.49 (m, 2H), 7.47-7.43 (m, 2H), 7.42-7.33 (m, 4H), 7.21-7.12 (m, 3H), 7.10 (s, 1H), 5.54, (s, 1H), 4.75 (s, 2H), 4.71 (s, 2H), 4.46-4.40 (m, 2H), 4.34-4.28 (m, 2H), 3.89-4.06 (m, 3H), 3.78-3.66(m, 2H), 3.27-3.03 (m, 2H), 2.32-2.20 (m, 2H), 2.18-2.08 (m, 2H), 2.02-1.91 (m, 2H), 1.75-1.86 (m, 2H).

LCMS (ESI): for C ₃₉ H ₃₆ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : calcd: 727.20, found: 727.20

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.76 (br s, 1H), 7.55 (br s, 1H), 7.53-7.47 (m, 2H), 7.48-7.44 (m, 2H), 7.42-7.36 (m, 3H), 7.34 (d, J = 8.4 Hz, 1H), 7.18-7.13 (m, 2H), 7.11 (dd, J = 6.9, 2.0 Hz, 2H), 4.73 (s, 2H), 4.71 ( s, 2H), 4.09-3.90 (m, 5H), 3.34-3.29 (m, 1H), 3.06-3.01 (m, 1H), 2.82-2.75 (m, 1H), 2.30-2.23 (m, 2H), 2.18-2.05 (m, 2H), 2.02-1.96 (m, 1H), 1.83-1.74 (m, 1H).LCMS(ESI): For C ₃₈ H ₃₂ Cl ₂ N ₄ O ₆ ; [M+H] ⁺ : Calculated: 711.18, Found: 711.51

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.76 (br s, 1H), 7.53-7.44 (m, 4H), 7.42-7.32 (m, 4H), 7.18-7.10(m, 4H), 4.74 ( s, 2H), 4.71 (s, 2H), 4.38-4.25 (m, 2H), 4.04-3.70 (m, 4H), 3.31-3.04 (m, 2H), 2.40-1.66 (m, 8H).

LCMS (ESI): for C ₃₈ H ₃₄ Cl ₂ N ₄ O ₅ ; [M+H] ⁺ : calculated: 697.20, found: 697.47. Example 14 PD-1/PD-L1 Homogeneous Time-Resolved Fluorescence (HTRF) Binding Analysis

The assay was performed in a standard black 384-well polystyrene plate with a final volume of 20 μL. The inhibitor was first serially diluted in DMSO and then added to the wells before the addition of other reaction components. The final concentration of DMSO in the assay was 1%. The assay was performed at 25°C using 0.05% Tween-20 and 0.1% BSA in PBS buffer (pH 7.4). Recombinant human PD-L1 protein (19-238) with a His tag at the C-terminus was purchased from AcroBiosystems (PD1-H5229). Recombinant human PD-1 protein (25-167) with an Fc tag at the C-terminus was also purchased from AcroBiosystems (PD1-H5257). PD-L1 and PD-1 proteins were diluted in assay buffer and 10 μL was added to the plate wells. The plates were centrifuged and the proteins were pre-incubated with inhibitors for 40 minutes. After incubation, 10 μL of HTRF detection buffer supplemented with cryptate-labeled anti-human IgG (Perkin Elmer-AD0212) specific for Fc and anti-His antibodies coupled to SureLight®-allophycocyanin (APC, PerkinElmer-AD0059H) was added. After centrifugation, the plates were incubated at 25°C for 60 minutes before reading on a PHERAstar FS plate reader (665 nm/620 nm ratio). The final concentrations in the assay were 3 nM PD1, 10 nM PD-L1, 1 nM anti-human IgG, and 20 nM anti-His allophycocyanin. _IC50 determinations were performed by fitting a curve of percent control activity versus the logarithm of inhibitor concentration. Table 1 Compound ID HTRF IC ₅₀ (nm) Compound ID HTRF IC ₅₀ (nm) Compound ID HTRF IC ₅₀ (nm) 281 26.74 454 1.42 523 10.89 303 187.43 457 8.78 527 9.41 338 1.46 458 1686.1 528 5.04 448 5.45 459 435.78 529 8.62 354 2184.69 460 9.9 530 76.88 361 1.75 461 4.06 531 7.22 363 2.16 465 3.58 533 7.11 372 3000 466 5 535 4.41 373 3.79 468 4.9 536 4.31 374 68.16 469 4.02 538 8.18 375 2707.11 470 2.28 539 7.73 376 199.54 471 4.7 540 10.5 378 69.66 472 5.27 542 5.24 381 145.86 475 3.23 545 4.81 382 3000 476 5.22 550 3.96 384 260.37 478 8.76 551 2.64 385 263.89 479 6.02 552 7.25 386 3000 480 5.48 554 5.51 388 1.1 481 4.69 557 5.33 394 470.58 482 6.72 558 10.64 395 6.26 483 6.31 559 8.98 396 11.66 484 6.53 560 7.78 397 304.73 486 1382.63 561 8.95 401 9.78 487 650.11 563 7.18 402 1.98 488 6.2 568 11.27 403 191.26 489 6.31 570 10.87 408 571.47 490 3.51 571 16.04 411 6.14 491 5.22 573 287.3 412 3000 492 3.15 576 10.94 413 7.05 493 5.59 585 10.87 414 737.11 494 6.95 588 7.35 415 103.69 500 10.69 590 6.04 422 3000 503 4.19 591 17.8 427 68.65 513 8.23 596 6.47 429 20.81 514 4.46 597 10.14 430 3000 515 8.71 607 13.89 445 5.01 516 10.5 608 28.1 449 3.89 517 7.46 614 7.7 450 6.47 518 7.19 Example 15 PD-L1 internalization 1. Experimental protocol

Day 1: Inoculation of cells: PD-L1/CHO-K1 cells were trypsinized in a flask, and then the cell number was counted and diluted to 1×10 ⁵ cells/ml. The cells were inoculated into a 6-well plate (Corning, #3516), 2 mL/well. The plate was incubated in a 37°C, 5% CO ₂ incubator for 24 hours.

Day 2: Prepare compounds and treated cells: Dilute GLC01-258 from 15 mM to 0.5 mM with DMSO and serially dilute 15 mM compound from 15 mM to 15 nM with DMSO, then dilute compound 500 times with assay buffer. Prepare 0.2% DMSO in assay buffer for vehicle control and low control. Remove plate, aspirate medium and discard. Add 2 mL of diluted compound, vehicle control, and low control to the corresponding wells. Incubate plate at 37°C, 5% CO ₂ incubator for 17 hours.

Day 3: Prepare samples for FACS: After 17 hours of incubation, discard the medium and wash with PBS. Digest the cells in each well with trypsin. Centrifuge and discard the supernatant, then wash the cells twice with DPBS (without Ca ²⁺ , Mg ²⁺ ). Dilute the antibody (PE-conjugated mouse anti-human CD274) tenfold with DPBS, and then add the staining solution to the compound-treated samples and the vehicle control samples. The low control only adds DPBS without antibody. Incubate the plate at room temperature for 20 minutes and protect from light. After 20 minutes, wash the samples twice with DPBS. Then centrifuge and discard the supernatant. Resuspend the cells in 300 uL DPBS and transfer the samples to 5 mL polystyrene round-bottom tubes (Falcon, #352054) and then analyze them using BD FACSCanto. 2. Data Analysis

The PD-L1 signal of the vector control was set to 100%, and the PD-L1 signal of the low control was set to 0%. The PD-L1 signal of the compound-treated samples was then calculated.

0% PD-L1 signal: low control stained without anti-CD274

100% PD-L1 signal: vector control stained with anti-CD274

Activation % of PD-L1 internalization = 1 - PD-L1 signal of the compound. Table 2 Compound ID Internalization Compound ID Internalization Compound ID Internalization 338 6.80% 481 8.70% 551 3.10% 363 5.50% 483 19.80% 552 4.90% 373 3.20% 500 19.50% 553 9.40% 388 6.10% 503 3.50% 554 23.20% 395 2.50% 517 22.80% 557 20.30% 402 2.40% 523 73.90% 560 3.10% 411 66.90% 528 13.30% 561 5.30% 445 20.50% 529 98.10% 563 25.50% 449 12.20% 530 1.60% 585 39.00% 454 5.20% 533 32.70% 588 2.00% 460 1.90% 535 21.00% 590 3.50% 465 67.00% 536 7.90% 596 2.90% 468 20.90% 538 3.60% 608 2.60% 470 20.90% 539 11.60% 613 4.80% 472 17.20% 540 10.60% 614 45.70% 475 31.50% 542 3.30% 620 43.90% 476 6.10% 545 7.80% 480 5.70% 550 3.00% Example 16 PD-L1 dimerization

Compounds were tested in a biochemical protein-protein interaction assay to determine whether they could specifically dimerize the extracellular domain of PD-L1.

(1) For 10+0 pts, serially dilute cpd 1:3 in DMSO for each column (refer to dilution plate diagram)

(2) Use Echo to transfer 0.2 μL of the cpd solution in each row to a 384-well plate, with each column containing 2 replicates (see plate map).

(3) Add 20 μL of the prepared mixture including PDL1-Eu and PDL1-A2 solutions to the assay plate and centrifuge at 1000 rpm for 1 min.

(4) Incubate at 25°C for 120 min.

(5) Read the fluorescent signal on the Envision 2104 reader.

(6) Read the ratiometric (665 nm)/615 nm) signal on the Envision.

(7) The raw data were analyzed using equation (V. Data Analysis). Table 3 Compound ID Dimerization EC ₅₀ (nm) Compound ID Dimerization EC ₅₀ (nm) Compound ID Dimerization EC ₅₀ (nm) 411 356.8 535 354.3 596 121.3 454 326.4 550 224.7 614 50.36 465 77.9 554 55.82 620 61.6 472 358.6 563 37.71 641 58.11 523 1330 585 130.3 648 59.15 528 174.9 588 154.4 649 63.7 533 90.96 590 220.1 Example 17 PDL1 Jurkat-NFAT reporter assay a. Preparation of Hep3B-OS8-hPDL1

1. Hep3B-OS8-hPDL1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and 100 μg/mL of G418 and hygromycin B.

2. Resuspend the cells in RPMI 1640 medium containing 10% FBS and adjust the cell density to 1.25E5 cells/mL.

3. Inoculate cells into 96-well flat-bottom plates (1.25E4 cells/100 μL/well). b. Preparation of compound solution

4. Remove the medium from the pre-plated Hep3B-OS8-PDL1 cells and wash once with 200 μL of assay medium.

5. Prepare compound dilutions as designed in RPMI 1640 medium with 10% FBS.

6. Compounds were added to each well at 9 concentrations (3, 1, 0.3, 0.1, 0.03, 0.01, 0.003, 0.001, and 0.0003 µM) in a volume of 50 μL. Keytruda at a concentration of 5 μg/mL served as a positive control.

7. Incubate at 37°C, 5% CO ₂ for 20-30 minutes. c. Preparation of Jurkat-NFAT-PD1

8. Jurkat-NFAT-PD1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and 1000 μg/mL hygromycin B and 0.3 μg/mL puromycin.

9. On the second day of the assay, resuspend the cells in RPMI 1640 medium containing 10% FBS and adjust the cell density to 2.5E5 cells/mL.

10. Seed the cells into 96-well flat-bottom plates (1.25E4 cells/50 μL/well).

11. Incubate the assay plate in a humidified 37°C, 5% CO ₂ incubator for 6 hours.

12. Allow the cultured cells to equilibrate at room temperature for 5-10 minutes.

13. Add an equal volume (100 μL/well) of ONE-Glo ^™ Luciferase Assay System to each well and wait at least 3 minutes to allow complete cell lysis and measurement in the luminometer. Table 4 Compound ID Jurkat-NFAT EC ₅₀ (nm) 465 0.3 529 111.1 Example 18 a. Hep3B-OS8-hPDL1 and T cell co-culture assay for tumor preparation

1. Hep3B-OS8-hPDL1 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin, and 100 μg/mL of G418 and hygromycin B.

2. Hep3B-OS8-hPDL1 cells were harvested and treated with 10 µg/mL mitomycin C at 37°C for 1.5 hours, then the cells were washed extensively four times with PBS.

3. Resuspend the cells in RPMI 1640 medium containing 10% FBS and adjust the cell density to 5E5 cells/mL.

4. Inoculate cells into 96-well flat-bottom plates (2.5E4 cells/50 μL/well). b. CD3+ T cell isolation (30 mL blood )

5. Dilute the human blood sample from an individual donor with an equal volume of sterile PBS, e.g., add 25 mL of sterile PBS to 25 mL of fresh whole blood, and mix thoroughly by gentle shaking.

6. Transfer 15 mL of Lymphoprep medium to a new 50 mL centrifuge tube.

7. Add the diluted blood sample as gently as possible onto the surface of the Ficoll medium to ensure that there is a clear boundary between the two liquids and that the volume ratio of Ficoll to diluted blood (30 mL) is 1:2.

8. During centrifugation, gently move the tube to the centrifuge with acceleration (5) and minimum deceleration (0) settings at 1000 × g, 25 min, 20°C.

9. After centrifugation, four interfaces can be observed, from top to bottom, they are plasma layer, monocyte layer, Ficoll medium layer and RBC layer, and move the tube as gently as possible to keep the four interfaces separated. Pipette and carefully transfer the second layer of monocytes to another new sterile centrifuge tube, and if it is unavoidable, pipette a certain volume of plasma instead of Ficoll medium.

10. Add three times the volume of PBMCs to the tube with PBMCs in sterile PBS.

11. Wash cells twice with 5 to 10 mL PBS before counting cells using a cell counter. Centrifuge at 350 × g, 10 min, 20°C. Use settings of acceleration (5) and deceleration (5) during centrifugation.

12. Resuspend the cells in the recommended medium and adjust the density of PBMCs to a final concentration of 5E7 cells/mL.

13. Isolate CD3+ T cells using EasySepTM Human T Cell Isolation Kit (STEMCELL Technologies #17951) and seed the cells into 96-well flat-bottom plates (5E4 cells/100 μL/well). c. Preparation of compound solution

14. Prepare compound dilutions in RPMI 1640 medium with 10% FBS as designed.

15. Add 50 μL of compound to each well. 7 concentrations (0.03, 0.1, 0.3, 1, 3, 10, and 30 µM) of 3 compounds (GLC01-258, GLC01-269, GLC01-465) and the same concentration (1 µM) of 6 compounds (GLC01-411, GLC01-292, GLC01-445, GLC01-475, GLC01-470, and GLC01-468)}

16. Keytruda was used as a positive control at a concentration of 5 μg/mL.

17. Incubate at 37°C, 5% _CO2 for 72 hours.

18. Collect the supernatant by centrifugation and measure IFN-γ using ELISA. Table 5 Compound ID Tumor and T cell EC ₅₀ (nm) 465 1.2 529 324.9 533 6.032 554 5.1 Example 19 Mouse PK study

(1) The compound was weighed and dissolved in a 1 mg/mL 5% saline solution, shaken thoroughly, and sonicated to form a colorless transparent solution. After an overnight fast, the solution was orally administered to a group of 3 mice at a dose of 10 mg/kg.

(2) Blood was collected from the submandibular vein and sodium heparin was used for anticoagulation. Blood was placed on ice after collection and plasma was separated by centrifugation within 1 hour (centrifugation conditions: 8000 rpm, 6 minutes, 2-8°C). Blood sampling time points were 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours.

(3) Store the sample in a -20°C freezer. Add 160 μL of ice-cold acetonitrile containing internal standard to the plasma sample (40 μL), vortex for 3 min, and centrifuge at 11,000 rpm for 5 min. Add 100 μL of the supernatant to 100 μL of water and inject 5 μL of the supernatant into the LC/MS/MS instrument to detect the compound (if the compound is an ester, the acid is detected).

The data are in Table 6: Table 6 Compound ID Observed compounds C _max (ng/mL) _Tmax (h) AUC _last (h·ng/mL) T _1/2 (h) 527 528 312 0.333 326 1.04 537 465 1426 0.333 1280 1.11 541 465 1533 0.333 1367 1.04 566 465 311 0.500 520 2.48 567 529 ND ND ND ND 589 554 575 0.250 338 1.04 594 465 916 0.500 1307 1.47 633 614 473 0.500 507 ND 634 639 702 0.500 571 ND 631 641 2160 0.667 2419 ND 638 629 4210 0.750 4639 ND 619 620 5237 0.500 10077 ND 640 647 3447 0.667 4055 ND 650 563 409 0.667 542 ND 652 653 68.2 0.833 94.7 ND

Although various embodiments have been described above, it should be understood that such disclosure is presented by way of example only and is not restrictive. Therefore, the breadth and scope of the subject compositions and methods should not be limited by any of the above exemplary embodiments, but should only be limited according to the scope of the attached patent application and its equivalent.

The above description is intended to teach a person skilled in the art how to implement the present application, and it is not intended to describe in detail all those obvious modifications and variations thereof, which will become apparent to a person skilled in the art after reading the description. However, it is intended that all such obvious modifications and variations are included within the scope of the present application as defined by the appended patent claims. The patent claims are intended to cover components and steps in any order that is effective to meet the intended purpose unless the context clearly indicates otherwise.

(without)

(without)

Claims (26)

A compound of formula (I) or a pharmaceutically acceptable salt thereof, a stereoisomer or a mixture of stereoisomers:

wherein A and B are each independently selected from halogen, cyano, _-N3 , alkyl and substituted alkyl, amine, alkylamine, alkoxy, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl; _Z1 is ^-CR1 = or -N=; _Z2 is ^-CR2 =; _Z3 is ^-CR3 = or -N=; _Z4 is ^-CR4 = or -N=; _Z5 is ^-CR5 =; _Z6 is ^-CR6 = or -N=; ^R1 and R ^{R 4} are each independently -H, halogen, cyano, alkyl, cycloalkyl having 3 to 10 carbon atoms, substituted alkyl, alkenyl having 2 to 12 atoms, alkynyl having 2 to 12 atoms, aryl having 5 to 14 ring atoms, amine, alkylamine, alkoxy, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl; R ² and R ^{R 5} are each independently -H, halogen, cyano, alkyl, cycloalkyl having 3 to 10 carbon atoms, substituted alkyl, alkenyl having 2 to 12 atoms, alkynyl having 2 to 12 atoms, aryl having 5 to 14 ring atoms, amine, alkylamine, alkoxy, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl; R ³ and R R ^{7 , R 8 , -C(R 7 )(R 8 )-, -CR 9 ＝, -NR 10 , -O- or -S-; X 1} and X 2 are each independently -C(R 11 )(R 12 )-, -N＝, -NR 13 , -S- or -O-; R 7 , R 8 , -C(R ⁷ )(R ⁸ )-, -CR 9 ＝, -NR 10 , -O- or -S-; X ₁ and X ₂ are each independently -C(R ¹¹ )(R ¹² )-, ^-N ＝, -NR ¹³ , -S- or -O-; R ₇ , R ⁸ , -C(R 7 )(R 8 )-, -CR 9 ^＝ , -NR ¹⁰ _, -O- or -S-; R ⁹ , R ¹¹ and R ¹² are each independently -H, halogen, cyano, alkyl, cycloalkyl having 3 to 10 carbon atoms, substituted alkyl, alkenyl having 2 to 12 atoms, alkynyl having 2 to 12 atoms, aryl having 5 to 14 ring atoms, amine, alkylamine, alkoxy, wherein the alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl; R ¹⁰ and R ^{W1 and W2} are each independently -H, alkyl, cycloalkyl having 3 to 10 carbon atoms, substituted alkyl, alkenyl having 2 to 12 atoms, alkynyl having 2 to 12 atoms, aryl having 5 to 14 ring atoms, alkylamine, alkoxy, wherein the alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl; _L1 and _L2 are each alkyl, substituted alkyl or heteroatom chain containing m atoms between ring 3 and _W1 and between ring 6 and _W2 , wherein m=0, 1, 2, 3, 4, 5 or 6; when m is ₀ , _W1 or _W2 is directly connected to the corresponding nitrogen in ring 3 or ring 6, respectively; ... ₂ are each independently hydrogen, a five-membered heterocyclic ring or a substituted five-membered heterocyclic ring, a six-membered heterocyclic ring or a substituted six-membered heterocyclic ring, a carboxyalkyl group or a substituted carboxyalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an amino acid ester, an amino acid amide, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as claimed in claim 1, wherein A and B are each independently selected from halogen, alkyl and substituted alkyl, cyano and -N ₃ , wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl, Z ₁ is -CR ¹ = or -N =; Z ₂ is -CR ² =; Z ₃ is -CR ³ = or -N =; Z ₄ is -CR ⁴ = or -N =; Z ₅ is -CR ⁵ =; Z ₆ is -CR ⁶ = or -N =; R ¹ and R ⁴ are each independently -H, -F, -Cl or -CH ₃ ; R ² and R ^{R 5} is each independently -H, -Cl, -F, -CH ₃ or -NH ₂ ; R ³ and R ⁶ are each independently -H, -Cl, -F, -CH ₃ or -NH ₂ ; X ₁ and X ₂ are each independently -C(R ¹¹ )(R ¹² )-, -N＝, -NH-, -N(R ¹³ )- or -O-; Y ₁ and Y ₂ are each independently -CH ₂ -, -CH＝, -NH-, -O-, -C(R ⁷ )(R ⁸ )-; R ⁷ , R 8 , R 11 and R ¹² are each independently -H, -F, -Cl or -CH 3; L ₁ and L ₂ are each alkyl or substituted alkyl containing m carbon atoms, wherein m=0, 1, 2, 3, 4, _{5 or 6; when m is 0, W 1 or} ^W 2 is each independently -C(R ¹¹ )(R 12 )-, -N＝, -NH-, -N(R 13 )- or -O-; _W1 and _W2 are each independently hydrogen, a five-membered heterocyclic ring or a _substituted five-membered heterocyclic ring, a six-membered heterocyclic ring or a substituted six-membered heterocyclic ring, a carboxyalkyl group or a substituted carboxyalkyl group, a cyanoalkyl group or a substituted cyanoalkyl group, an aminoalkyl group or a substituted aminoalkyl group, a hydroxyalkyl group or a substituted hydroxyalkyl group, an amino acid, an amino acid ester, an amino acid amide, a non-natural amino acid, an ester of a non-natural amino acid, or an amide of a non-natural amino acid. The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 1, wherein the compound comprises a core structure selected from formula (II) to (XXIII):

The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 3, wherein _L1 and _L2 are each independently _C1 - _C3 alkyl. A compound as described in any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, a stereoisomer or a mixture of stereoisomers, wherein _W1 and _W2 are each independently a type I side chain, wherein the type I side chain is selected from:

,

,

,

, -C(O)ONa, -CN, -CH ₂ OH, and -CH ₂ NH ₂ . A compound as described in any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, a stereoisomer or a mixture of stereoisomers, wherein _W1 and _W2 are each independently a type II side chain, wherein the type II side chain of _W1 has the following general formula:

wherein R ¹⁴ and R ¹⁵ are independently -H, alkyl or substituted alkyl, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl, and wherein the type II side chain of W ₂ has the following general formula:

wherein R ¹⁶ and R ¹⁷ are independently -H, alkyl or substituted alkyl, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl. The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 6, wherein R ¹⁴ is one of the following:

R ¹⁵ is independently -H, alkyl or substituted alkyl, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl, wherein R ¹⁶ is one of the following:

,

,

and

, and R ¹⁷ is independently -H, alkyl or substituted alkyl, wherein alkyl is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl or cyclohexyl. The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 6, wherein R ¹⁴ and R ¹⁶ are each independently selected from:

The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 6, wherein _W1 or _W2 is L-serine. The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in claim 9, wherein _W1 and _W2 are both L-serine. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as described in claim 6, wherein _W1 or _W2 is an ester of L-serine. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as claimed in claim 11, wherein _W1 and _W2 are both esters of L-serine. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as described in claim 6, wherein _W1 and/or _W2 is an ester of L-serine. The compound or pharmaceutically acceptable salt thereof, stereoisomer or mixture of stereoisomers as described in any one of claims 1 to 4, wherein _W1 and _W2 are independently selected from:

The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

,

or

, and W ₂ is H, and L ₂ does not exist (i.e., m=0). The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

,

or

, _W2 is a type I or type II side chain. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

, _W2 is a type I or type II side chain. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

, _W2 is a type I or type II side chain. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

, _W2 is a type I or type II side chain. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof according to any one of claims 1 to 4, wherein _W1 is

, _W2 is

. The compound or pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as described in any one of claims 1 to 4, wherein _L1 and _L2 are each independently _C1 - _C3 alkyl. A compound or a pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as described in claim 1, wherein the compound is selected from the group consisting of compounds 281, 303, 448, 338, 354, 361, 363, 372, 376, 381, 382, 388, 395, 396, 401, 402, 457, 466, 481, 445, 449, 454, 460, 461, 465, 468, 469, 470, 471, 475, 476, 478, 482, 483, 484, 490, 491, 493, 513, 527, 528, 536, 537, 541, 545, 551, 552, 553, 560, 561, 563, 564, 565, 566, 567, 568, 569, 570, 571, 575, 576, 578, 579, 580, 581, 582, 583, 584, 585 66, 570, 571, 573, 588, 479, 480, 563, 550, 554, 589, 561, 557, 586, 585, 514, 373, 374, 375, 378, 384, 386, 427, 394, 397, 408, 411, 503, 530, 531, 53 5, 53 9, 540, 412, 413, 415, 459, 422, 430, 472, 567, 529, 558, 559, 538, 429, 385, 533, 516, 515, 542, 488, 489, 492, 450, 517, their pharmaceutically acceptable salts, their stereoisomers and mixtures of their stereoisomers. A compound or a pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof as described in claim 1, wherein the compound is selected from compounds 338, 361, 411, 465, 475, 483, 529, 533, 537, 541, 554, 585, 613, 614, 619, 620, 628, 633, 648, 649 in Table 1, pharmaceutically acceptable salts thereof, stereoisomers thereof and mixtures of stereoisomers thereof. A compound which is

or a pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof. Use of a compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer or mixture of stereoisomers thereof in the preparation of a drug for treating a disease or disease state associated with the interaction between PD-L1 and PD-1 in an individual. The use as described in claim 25, wherein the disease is cancer.